Bridges Lab Protocols - User contributions [en]

Mammary Gland Fat and SVF Separation

2021-01-25T19:51:55Z

Elhabbal:

Mammary Gland Fat and SVF Separation

* Warm PBS and the Erlenmeyer flask from the shaker in water bath at 37 degrees Celsius. Have an ice bucket ready.
* Dissect mouse MG. Be consistent with the side and the size of the MG dissected. Be careful that the dissection is clean without skin, fur, blood vessels, or unwanted tissue.
* Put the dissected MG in a petri dish with 1ml PBS to keep tissue moist.
* Dump out additional PBS while holding the MG with forceps to ensure it does not fall. This ensures that the tissue will not become too soggy and hinder proper mincing.
* Mince the tissue in the petri dish while maintaining a 30-45 degree angle. Mince using the bigger scissors for about 5-10 minutes until all the tissue chunks look smaller than 1mm.
* Cut the tip of a 1000uL pipette’s tip. Using this tip, transfer out the minced tissue from the petri dish into a 50mL tube.
* Add 1mL collagenase mix (BAT digestion mix) into the 50mL tube.
* Seal the tube lid with parafilm. Place in the Erlenmeyer flask in the shaker at 37 degrees Celsius and 300 strokes for about 45-50 minutes or until the tissue looks digested and the pieces are not big anymore. Check the tube every 5-10 minutes to ensure digestion.
* Once digestion in the shaker is over, pipette out the mixture into a 1.5mL tube. If the tissue is still big, cut the pipette tip’s tip to be able to pipette the tissue.
* Centrifuge the 1.5mL tube at room temperature (24 degrees Celsius) for 5 minutes at 500 rcf/gauge.
* Once centrifuging is done, the upper fat layer will be evident along with the chunky SVF lower layer as the pellet. The in-between layer solution is called the internatant.
* Cut a 200mL pipette tip’s tip and pipette out the fat layer into a clean 1.5mL tube. Record the volume removed and then split the fat equally into 2 1.5mL tubes. For example, if you remove 800uL of fat (with internatant), then you will end up with 2 1.5mL tubes that have 400uL fat each. Put the fat tubes on ice.
Note: It is hard to isolate the fat while pipetting, so you should anticipate to pipette out some internatant with the fat.
* Using a pipette, aspirate and dump the remaining internatant solution. Be careful not to pipette out the SVF pellet.
* Add 600uL of PBS to the pellet tube and mix vigorously using the pipette (by pipetting repeatedly) to break down the pellet. Pipette 300mL SVF+PBS solution into a 1.5mL tube. Pipette 300mL SVF+PBs again into another 1.5mL tube. Put the SVF tubes on ice.
* Dump the initial tube with the remaining SVF+PBS.
* Freeze the 2 fat tubes and the 2 SVF tubes in the -80 freezer for later use.

Mammary Gland Fat and SVF Separation

2020-12-22T21:53:15Z

Elhabbal: Created mammary gland fat and SVF separation protocol

Mammary Gland Fat and SVF Separation

* Warm PBS and the Erlenmeyer flask from the shaker in water bath at 37 degrees Celsius. Have an ice bucket ready.
* Dissect mouse MG. Be consistent with the side and the size of the MG dissected. Be careful that the dissection is clean without skin, fur, blood vessels, or unwanted tissue.
* Put the dissected MG in a petri dish with 1ml PBS to keep tissue moist.
* Dump out additional PBS while holding the MG with forceps to ensure it does not fall. This ensures that the tissue will not become too soggy and hinder proper mincing.
* Mince the tissue in the petri dish while maintaining a 30-45 degree angle. Mince using the bigger scissors for about 5-10 minutes until all the tissue chunks look smaller than 1mm.
* Cut the tip of a 1000uL pipette’s tip. Using this tip, transfer out the minced tissue from the petri dish into a 50mL tube.
* Add 1mL collagenase mix (BAT digestion mix) into the 50mL tube.
* Seal the tube lid with parafilm. Place in the Erlenmeyer flask in the shaker at 37 degrees Celsius and 300 strokes for about 45-50 minutes or until the tissue looks digested and the pieces are not big anymore. Check the tube every 5-10 minutes to ensure digestion.
* Once digestion in the shaker is over, pipette out the mixture into a 1.5mL tube. If the tissue is still big, cut the pipette tip’s tip to be able to pipette the tissue.
* Centrifuge the 1.5mL tube at room temperature (24 degrees Celsius) for 5 minutes at 500 rcf/gauge.
* Once centrifuging is done, the upper fat layer will be evident along with the chunky SVF lower layer as the pellet. The in-between layer solution is called the internatant.
Cut a 200mL pipette tip’s tip and pipette out the fat layer into a clean 1.5mL tube. Record the volume removed and then split the fat equally into 2 1.5mL tubes. For example, if you remove 800uL of fat (with internatant), then you will end up with 2 1.5mL tubes that have 400uL fat each. Put the fat tubes on ice.
Note: It is hard to isolate the fat while pipetting, so you should anticipate to pipette out some internatant with the fat.
* Using a pipette, aspirate and dump the remaining internatant solution. Be careful not to pipette out the SVF pellet.
* Add 600uL of PBS to the pellet tube and mix vigorously using the pipette (by pipetting repeatedly) to break down the pellet. Pipette 300mL SVF+PBS solution into a 1.5mL tube. Pipette 300mL SVF+PBs again into another 1.5mL tube. Put the SVF tubes on ice.
* Dump the initial tube with the remaining SVF+PBS.
* Freeze the 2 fat tubes and the 2 SVF tubes in the -80 freezer for later use.

Weigh-Suckle-Weigh Milk Volume Measurement

2020-12-22T21:38:15Z

Elhabbal:

Weigh-Suckle-Weigh Protocol at PND10.5

* This is usually done at PND10.5 (Usually denote PND0.5 as the day pups were born).
* Weigh dam alone using the regular metric weighing scale (in the procedure room) and weigh aggregate of pups by placing all pups on weighing paper using the analytical scale (in the lab).
* Keep the dam in the original cage with food and water bottle ad-lib. Place pups in a new cage without food or water, but keep the normal cage enrichment (the bedding pouch, tear it open, and put it on top of pups as a blanket).
* Put the pup cage on a heating pad (set heat to “high”) so they will remain warm for the separation period. Put ¾ of the cage on the pad and leave a ¼ on the silver bench so pups can choose to move around if they get too warm. I usually place the dam’s cage next to the pups with the similar ¾ ¼ alignment on the heating pad and bench, respectively.
* Keep the pups and dam separated for two hours.
* After two hours of separation, weigh the pup aggregate weight on the analytical scale and weigh the dam using the metric weighing scale.
* Recombine the pups and dam in the original cage.
* After one hour of nursing, weigh the dam again using the metric weighing scale then weigh the aggregate of pups using the analytical scale.

Note 1:
* The data collected should look something like this for the dam and pups for all time points. This is an example to highlight the 3 data points for weight measurements.
Dam wt 26.9g ---> after 2 hour separation 25.7g ---> after 1 hour nursing 25.0

Pups wt 22.71 ---> after 2 hour separation 22.55 ---> after 2 hours nursing 23.20g

Note 2:
* The weight of the dam may decrease after 2-hour separation from pups, do not worry about that (despite assuming the dam should retain more milk and gain weight).

Weigh-Suckle-Weigh Milk Volume Measurement

2020-12-22T21:37:46Z

Elhabbal: Weigh-suckle-weigh protocol to measure milk output per dam at PND10.5 protocol created

Weigh-Suckle-Weigh Protocol at PND10.5

* This is usually done at PND10.5 (Usually denote PND0.5 as the day pups were born).
* Weigh dam alone using the regular metric weighing scale (in the procedure room) and weigh aggregate of pups by placing all pups on weighing paper using the analytical scale (in the lab).
* Keep the dam in the original cage with food and water bottle ad-lib. Place pups in a new cage without food or water, but keep the normal cage enrichment (the bedding pouch, tear it open, and put it on top of pups as a blanket).
* Put the pup cage on a heating pad (set heat to “high”) so they will remain warm for the separation period. Put ¾ of the cage on the pad and leave a ¼ on the silver bench so pups can choose to move around if they get too warm. I usually place the dam’s cage next to the pups with the similar ¾ ¼ alignment on the heating pad and bench, respectively.
* Keep the pups and dam separated for two hours.
* After two hours of separation, weigh the pup aggregate weight on the analytical scale and weigh the dam using the metric weighing scale.
* Recombine the pups and dam in the original cage.
* After one hour of nursing, weigh the dam again using the metric weighing scale then weigh the aggregate of pups using the analytical scale.

Note 1:
* The data collected should look something like this for the dam and pups for all time points. This is an example to highlight the 3 data points for weight measurements.
Dam wt 26.9g ---> after 2 hour separation 25.7g ---> after 1 hour nursing 25.0
Pups wt 22.71 ---> after 2 hour separation 22.55 ---> after 2 hours nursing 23.20g

Note 2:
* The weight of the dam may decrease after 2-hour separation from pups, do not worry about that (despite assuming the dam should retain more milk and gain weight).

Weigh-Suckle-Weigh Milk Volume Measurement

2020-12-22T21:34:36Z

Elhabbal: Created page with "Weigh-Suckle-Weigh Protocol at PND10.5 1. This is usually done at PND10.5 (Usually denote PND0.5 as the day pups were born). 2. Weigh dam alone using the regular metric weigh..."

Weigh-Suckle-Weigh Protocol at PND10.5

1. This is usually done at PND10.5 (Usually denote PND0.5 as the day pups were born).
2. Weigh dam alone using the regular metric weighing scale (in the procedure room) and weigh aggregate of pups by placing all pups on weighing paper using the analytical scale (in the lab).
3. Keep the dam in the original cage with food and water bottle ad-lib. Place pups in a new cage without food or water, but keep the normal cage enrichment (the bedding pouch, tear it open, and put it on top of pups as a blanket).
4. Put the pup cage on a heating pad (set heat to “high”) so they will remain warm for the separation period. Put ¾ of the cage on the pad and leave a ¼ on the silver bench so pups can choose to move around if they get too warm. I usually place the dam’s cage next to the pups with the similar ¾ ¼ alignment on the heating pad and bench, respectively.
5. Keep the pups and dam separated for two hours.
6. After two hours of separation, weigh the pup aggregate weight on the analytical scale and weigh the dam using the metric weighing scale.
7. Recombine the pups and dam in the original cage.
8. After one hour of nursing, weigh the dam again using the metric weighing scale then weigh the aggregate of pups using the analytical scale.
Notes: The data collected should look something like this for the dams and pups at each time point.
Dam wt 26.9g ---> after 2 hour separation 25.7g ---> after 1 hour nursing 25.0
Pups wt 22.71 ---> after 2 hour separation 22.55 --->after 2 hours nursing 23.20g

Notes: The weight of the dam may decrease after 2-hour separation from pups, do not worry about that (despite assuming the dam should retain more milk and gain weight).

Dexamethasone Treatment of Mice

2019-12-30T20:26:14Z

Elhabbal: /* Calculation */

[[Category:Mouse Work]]
[[Category:Animal Studies]]
[[Category:Drug Treatment]]
[[Category:Insulin Sensitivity]]
[[Category:Insulin Signaling]]
[[Category:Metabolism]]

We use water-soluble Dex from Sigma (D2915). It says on the bottle how many mg/g is actually Dex. The calculations below are for a bottle where the powder was 65mg/g. Dex

==For i.p. injection 5 mg/kg==

* Makes 15mL stock solution of 1mg/mL
* dissolve 0.231g Dex powder in 15mL PBS

<pre>BW (g) X 0.005 = injection volume (mL)</pre>

===Calculation===
<pre>
65mg dex/ = 15mg dex/ = 0.231g
1000mg powder ? mg powder
</pre>

==For drinking water administration ~1mg/kg/day==

* Makes 1L of drinking water
* dissolve 0.092g in 1L water

===Calculation===
* 1mg Dex/kg/day because mice drink ~5mL /day so want 0.2 mg Dex/kg/mL
* average mouse weighs 0.03kg so want 0.006mg Dex/mL or 6mg/L
<pre>
65mg dex/ = 6mg dex/ => X= 0.092g solid powder
1000mg solid powder ? (X) mg solid powder
</pre>
*This assumes that the mice are about 30g and they drink 5mL/day.

Dexamethasone Treatment of Mice

2019-12-30T20:25:53Z

Elhabbal: /* Calculation */

[[Category:Mouse Work]]
[[Category:Animal Studies]]
[[Category:Drug Treatment]]
[[Category:Insulin Sensitivity]]
[[Category:Insulin Signaling]]
[[Category:Metabolism]]

We use water-soluble Dex from Sigma (D2915). It says on the bottle how many mg/g is actually Dex. The calculations below are for a bottle where the powder was 65mg/g. Dex

==For i.p. injection 5 mg/kg==

* Makes 15mL stock solution of 1mg/mL
* dissolve 0.231g Dex powder in 15mL PBS

<pre>BW (g) X 0.005 = injection volume (mL)</pre>

===Calculation===
<pre>
65mg dex/ = 15mg dex/ = 0.231g
1000mg powder ? mg powder
</pre>

==For drinking water administration ~1mg/kg/day==

* Makes 1L of drinking water
* dissolve 0.092g in 1L water

===Calculation===
* 1mg Dex/kg/day because mice drink ~5mL /day so want 0.2 mg Dex/kg/mL
* average mouse weighs 0.03kg so want 0.006mg Dex/mL or 6mg/L
<pre>
65mg dex/ = 6mg dex/ => X= 0.092g
1000mg solid powder ? (X) mg solid powder
</pre>
*This assumes that the mice are about 30g and they drink 5mL/day.

Dexamethasone Treatment of Mice

2019-12-30T20:25:23Z

Elhabbal: /* Calculation */

[[Category:Mouse Work]]
[[Category:Animal Studies]]
[[Category:Drug Treatment]]
[[Category:Insulin Sensitivity]]
[[Category:Insulin Signaling]]
[[Category:Metabolism]]

We use water-soluble Dex from Sigma (D2915). It says on the bottle how many mg/g is actually Dex. The calculations below are for a bottle where the powder was 65mg/g. Dex

==For i.p. injection 5 mg/kg==

* Makes 15mL stock solution of 1mg/mL
* dissolve 0.231g Dex powder in 15mL PBS

<pre>BW (g) X 0.005 = injection volume (mL)</pre>

===Calculation===
<pre>
65mg dex/ = 15mg dex/ = 0.231g
1000mg powder ? mg powder
</pre>

==For drinking water administration ~1mg/kg/day==

* Makes 1L of drinking water
* dissolve 0.092g in 1L water

===Calculation===
* 1mg Dex/kg/day because mice drink ~5mL /day so want 0.2 mg Dex/kg/mL
* average mouse weighs 0.03kg so want 0.006mg Dex/mL or 6mg/L
<pre>
65mg dex/ = 6mg dex/ => X= 0.092g
1000mg solid powder ? (X) mg powder
</pre>
*This assumes that the mice are about 30g and they drink 5mL/day.

Dexamethasone Treatment of Mice

2019-12-30T20:25:06Z

Elhabbal: /* Calculation */

[[Category:Mouse Work]]
[[Category:Animal Studies]]
[[Category:Drug Treatment]]
[[Category:Insulin Sensitivity]]
[[Category:Insulin Signaling]]
[[Category:Metabolism]]

We use water-soluble Dex from Sigma (D2915). It says on the bottle how many mg/g is actually Dex. The calculations below are for a bottle where the powder was 65mg/g. Dex

==For i.p. injection 5 mg/kg==

* Makes 15mL stock solution of 1mg/mL
* dissolve 0.231g Dex powder in 15mL PBS

<pre>BW (g) X 0.005 = injection volume (mL)</pre>

===Calculation===
<pre>
65mg dex/ = 15mg dex/ = 0.231g
1000mg powder ? mg powder
</pre>

==For drinking water administration ~1mg/kg/day==

* Makes 1L of drinking water
* dissolve 0.092g in 1L water

===Calculation===
* 1mg Dex/kg/day because mice drink ~5mL /day so want 0.2 mg Dex/kg/mL
* average mouse weighs 0.03kg so want 0.006mg Dex/mL or 6mg/L
<pre>
65mg dex/ = 6mg dex/ => X= 0.092g
1000mg solid powder ? (X) mg powder
</pre>
*This assumes that the mice are about 30g and they drink 5mL/day.

SOP - Bleach

2019-10-07T15:25:09Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
This standard operating procedure outlines the handling and use of bleach. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling bleach.
This document is modified from http://www.oseh.umich.edu/docs/SOP/Bleach.docx

Bleach is a general term that typically refers to an aqueous mixture that has a primary active ingredient of either Sodium Hypochlorite or Calcium Hypochlorite and usually at concentrations of 3 - 10%. Bleach is used primarily as an oxidizing, cleaning, or a bleaching agent, as a disinfectant or within drinking water and waste water purification systems.

Synonyms of bleach include sodium hypochlorite, Clorox, liquid bleach, antiformin, chlorox, Carrel-dakin solution, Chloros, Dakin’s solution, hychlorite, Javelle water.

==Useful Bleach Links==
* http://www.atsdr.cdc.gov/toxfaqs/tfacts184.pdf
* http://www.atsdr.cdc.gov/Mhmi/mmg184.pdf
* http://www.asc.co.id/uplimg/File/03%20zMSDS_NaClO_ASC%20R3.pdf
* http://www.cl2.com/documents/Incompatibility%20Chart.pdf

==Potential Hazards==
* Corrosive. May cause severe irritation or damage to eyes or skin. Vapor may irritate eyes and respiratory tract.
* Incompatible with many chemicals/agents. Hazardous gases (including chlorine and chloramines) may be released if bleach is mixed with an incompatible material.
* Exposure to these gases can cause coughing, shortness of breath, irritation to, or burning of the eyes, nose, and throat, chest pain, wheezing, fluid in the lungs, and nausea. Chlorine can also be absorbed through the skin and cause pain, inflammation, swelling, and blistering.
* Strong oxidizer. May initiate or promote combustion in other materials.

==Occupational Exposure Limits (OELs)==
* MIOSHA: 0.5 ppm Chlorine (from sodium hypochlorite), 8-hour PEL
* MIOSHA: 1 ppm Chlorine (from sodium hypochlorite), 15-minute STEL
* AIHA (WEEL): 2 mg/m3 Sodium Hypochlorite, 15-minute STEL

==Engineering Controls==
* An eyewash/drench hose combination unit must be available in the immediate work area for any work with corrosive materials, including bleach.
* If large quantities will be used, a safety shower will also be necessary. Contact OSEH at (734) 647-1143 for a determination of the need for a safety shower if there is not one available.
* Depending on the material’s pH or based on its ability to cause severe tissue damage, e.g., formaldehyde, methylene chloride, methyl ethyl ketone peroxide (MEKP), phenol, etc., the location of the emergency shower and/or emergency eyewash shall be within 25 to 100 feet from the hazardous operation.
* A system of local and/or general exhaust is recommended to keep employee exposures below MIOSHA Permissible Exposure Limits (PEL). Local exhaust ventilation (LEV) is generally preferred because it can control the emissions of the contaminant at its source, preventing dispersion of it into the general work area. Contact OSEH at (734) 647-1143 for a determination of the need for an LEV system, if there is not one available.

==Work Practice Controls==
* ALWAYS review the SDS of both bleach and any chemicals/agents before mixing them with bleach to ensure compatibility. A general list of incompatible materials can be found here.
* NEVER mix bleach with an unknown liquid or unknown residue.
* Do not mix bleach with any compound that is incompatible with oxidizers.
* Purchase bleach in the smallest containers that are practical for lab use.
* Work with the smallest practicable amount and lowest practicable concentration.

==Personal Protective Equipment (PPE)==
In order to select the appropriate PPE for the workplace, a Hazard Assessment is conducted. The hazard assessment determines the hazards and potential hazards associated with a task, machinery, or process. The appropriate PPE for the situation may be subsequently determined. The Hazard Assessment Form may be found here. It may be completed either by the workplace supervisor or the department’s OSEH representative.
* Personal protective equipment is especially important. Wear a buttoned lab coat, safety glasses (that meet the requirements of ANSI/ISEA Z87.1) or goggles if splashing may occur, and gloves for any work with bleach. Depending on the quantities and concentrations used, a face shield, impenetrable, chemical apron and sleeves (or coverall), and special gloves may be recommended.
* Should adequate dilution ventilation or LEV be present, respiratory protection should not be necessary. However, in the absence of these and when MIOSHA PEL, or other published occupational exposures limits (OEL) are anticipated to be exceeded, respiratory protection may be necessary. Contact OSEH at (734) 647-1143 for a determination of the need for a respirator.

==Transportation and Storage==
* Transport concentrated bleach solutions in secondary containment, preferably a polyethylene or other non-reactive acid/solvent bottle carrier.
* Store in well-ventilated areas with secondary containment, such as a non-reactive plastic bin.
* Store below eye level.
* Store away from metal (unless the metal has a corrosion-proof coating), and do not store under the sink.
* Store away from incompatibles and flammable materials. Always review the SDS of other chemicals in the storage area for compatibility with bleach.
* Avoid storing on the floor. If storing on the floor is necessary, use secondary containment.

==Waste Disposal==
If the bleach solution is at a household concentration or less, i.e., a maximum 10% concentration of sodium hypochlorite, and was used for disinfecting, limit discharges down a laboratory sink to less than 1-gallon (approx. 3.8 L). If the concentration was >10% and was used for disinfecting, limit the sink disposal to less than 2-cups (approx. 500 mL) of bleach solution. Also flush the drain with adequate volumes of water.

This does not apply to “waste” bleach, e.g., unused or expired bleach. Any leftover, unused product or expired bleach not used as intended by the manufacturer, or used as a disinfectant, needs be collected by OSEH Hazardous Materials Management (HMM) for proper disposal. Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, and waste collection. Also refer to the OSEH Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==
If the employee is in need of emergency medical attention, call 911 immediately.

In general, flush affected eyes or skin with water for at least 15 minutes, then seek medical attention (see below).
* Eyes: In case of contact, immediately flush eyes with plenty of water for at least 15 minutes. Get medical aid.
* Skin: In case of contact, immediately flush skin with plenty of water. Remove contaminated clothing and shoes. Get medical aid. Wash clothing before reuse.
* Ingestion: Get medical aid immediately. Do not induce vomiting unless directed to do so by medical personnel. Never give anything by mouth to an unconscious person. If vomiting occurs naturally, have victim lean forward. Wash mouth with water, and then give plenty of milk or water to drink and obtain urgent medical attention.
* Inhalation: If inhaled, remove to fresh air. If not breathing, give artificial respiration. Get medical aid.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==Treatment Facilities==
* U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
* University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
* UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

==Spill Procedures==
* When a spill occurs, personal safety should always come first.
* Alert and clear everyone in the immediate area where the spill occurred.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., less than 1 Gallon or 3.5 Liters. A major/large chemical spill requires active assistance from emergency personnel.

For minor (small) spills
* Use proper personal protective equipment as indicated above.
* Absorb the spilled bleach using an absorbent, non-combustible material such as appropriate sorbent pads, sand, or vermiculite. Do not use combustible materials such as sawdust.
* Collect residue, place in container and contact OSEH-HMM (734) 763-4568 for proper disposal.

For major (large) spills
Report large chemical spills greater than 1 gallon or 3.5 liters in corridors or common areas, e.g., hallways, elevators, eating areas, rest rooms, offices, etc., to University of Michigan Division of Public Safety and Security (DPSS) by calling 911.

Additional Spill Response Steps:

===Minor Chemical Spill===
* Alert people in immediate area of spill.
* Open outside windows, if possible.
* Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.
* Avoid breathing vapors from spill.
* Confine spill to as small an area as possible.
* Do not wash spill down the drain.
* Use appropriate spill kits/sorbents to neutralize corrosives and/or absorb spill. Collect contaminated materials and residues and place in container. For powdered chemicals sweep carefully to avoid generation of dust or, if appropriate, use moist sorbent pads or wet the powder with a suitable solvent and then wipe with a dry cloth. Contact OSEH-HMM (734) 763-4568 for proper disposal.
* Clean spill area with water.

===Major Chemical Spill===
* Attend to injured or contaminated persons and remove them from exposure.
* Alert people in the laboratory to evacuate.
* Close doors to affected area.
* Post warnings to keep people from entering the area.
* Have person available that has knowledge of incident and laboratory to assist emergency personnel.

===Additional Spill Links===
* http://www.oseh.umich.edu/pdf/chemspil.pdf
* http://www.oseh.umich.edu/emer-chemical.shtml

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling bleach.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 16:42, 12 October 2016 (UTC)
* [[User:Ccousz|Ccousz]] ([[User talk:Ccousz|talk]])
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:17, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 18:47, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:45, 12 June 2017 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 14:52, 13 June 2017 (UTC)
*[[User:Nlambrec|Nlambrec]] ([[User talk:Nlambrec|talk]]) 21:08, 17 February 2019 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])

Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☒ NO

Laboratory Director - Dave Bridges Revision Date - 2016-07-26

SOP - Bleach

2019-10-07T15:24:37Z

Elhabbal:

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
This standard operating procedure outlines the handling and use of bleach. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling bleach.
This document is modified from http://www.oseh.umich.edu/docs/SOP/Bleach.docx

Bleach is a general term that typically refers to an aqueous mixture that has a primary active ingredient of either Sodium Hypochlorite or Calcium Hypochlorite and usually at concentrations of 3 - 10%. Bleach is used primarily as an oxidizing, cleaning, or a bleaching agent, as a disinfectant or within drinking water and waste water purification systems.

Synonyms of bleach include sodium hypochlorite, Clorox, liquid bleach, antiformin, chlorox, Carrel-dakin solution, Chloros, Dakin’s solution, hychlorite, Javelle water.

==Useful Bleach Links==
* http://www.atsdr.cdc.gov/toxfaqs/tfacts184.pdf
* http://www.atsdr.cdc.gov/Mhmi/mmg184.pdf
* http://www.asc.co.id/uplimg/File/03%20zMSDS_NaClO_ASC%20R3.pdf
* http://www.cl2.com/documents/Incompatibility%20Chart.pdf

==Potential Hazards==
* Corrosive. May cause severe irritation or damage to eyes or skin. Vapor may irritate eyes and respiratory tract.
* Incompatible with many chemicals/agents. Hazardous gases (including chlorine and chloramines) may be released if bleach is mixed with an incompatible material.
* Exposure to these gases can cause coughing, shortness of breath, irritation to, or burning of the eyes, nose, and throat, chest pain, wheezing, fluid in the lungs, and nausea. Chlorine can also be absorbed through the skin and cause pain, inflammation, swelling, and blistering.
* Strong oxidizer. May initiate or promote combustion in other materials.

==Occupational Exposure Limits (OELs)==
* MIOSHA: 0.5 ppm Chlorine (from sodium hypochlorite), 8-hour PEL
* MIOSHA: 1 ppm Chlorine (from sodium hypochlorite), 15-minute STEL
* AIHA (WEEL): 2 mg/m3 Sodium Hypochlorite, 15-minute STEL

==Engineering Controls==
* An eyewash/drench hose combination unit must be available in the immediate work area for any work with corrosive materials, including bleach.
* If large quantities will be used, a safety shower will also be necessary. Contact OSEH at (734) 647-1143 for a determination of the need for a safety shower if there is not one available.
* Depending on the material’s pH or based on its ability to cause severe tissue damage, e.g., formaldehyde, methylene chloride, methyl ethyl ketone peroxide (MEKP), phenol, etc., the location of the emergency shower and/or emergency eyewash shall be within 25 to 100 feet from the hazardous operation.
* A system of local and/or general exhaust is recommended to keep employee exposures below MIOSHA Permissible Exposure Limits (PEL). Local exhaust ventilation (LEV) is generally preferred because it can control the emissions of the contaminant at its source, preventing dispersion of it into the general work area. Contact OSEH at (734) 647-1143 for a determination of the need for an LEV system, if there is not one available.

==Work Practice Controls==
* ALWAYS review the SDS of both bleach and any chemicals/agents before mixing them with bleach to ensure compatibility. A general list of incompatible materials can be found here.
* NEVER mix bleach with an unknown liquid or unknown residue.
* Do not mix bleach with any compound that is incompatible with oxidizers.
* Purchase bleach in the smallest containers that are practical for lab use.
* Work with the smallest practicable amount and lowest practicable concentration.

==Personal Protective Equipment (PPE)==
In order to select the appropriate PPE for the workplace, a Hazard Assessment is conducted. The hazard assessment determines the hazards and potential hazards associated with a task, machinery, or process. The appropriate PPE for the situation may be subsequently determined. The Hazard Assessment Form may be found here. It may be completed either by the workplace supervisor or the department’s OSEH representative.
* Personal protective equipment is especially important. Wear a buttoned lab coat, safety glasses (that meet the requirements of ANSI/ISEA Z87.1) or goggles if splashing may occur, and gloves for any work with bleach. Depending on the quantities and concentrations used, a face shield, impenetrable, chemical apron and sleeves (or coverall), and special gloves may be recommended.
* Should adequate dilution ventilation or LEV be present, respiratory protection should not be necessary. However, in the absence of these and when MIOSHA PEL, or other published occupational exposures limits (OEL) are anticipated to be exceeded, respiratory protection may be necessary. Contact OSEH at (734) 647-1143 for a determination of the need for a respirator.

==Transportation and Storage==
* Transport concentrated bleach solutions in secondary containment, preferably a polyethylene or other non-reactive acid/solvent bottle carrier.
* Store in well-ventilated areas with secondary containment, such as a non-reactive plastic bin.
* Store below eye level.
* Store away from metal (unless the metal has a corrosion-proof coating), and do not store under the sink.
* Store away from incompatibles and flammable materials. Always review the SDS of other chemicals in the storage area for compatibility with bleach.
* Avoid storing on the floor. If storing on the floor is necessary, use secondary containment.

==Waste Disposal==
If the bleach solution is at a household concentration or less, i.e., a maximum 10% concentration of sodium hypochlorite, and was used for disinfecting, limit discharges down a laboratory sink to less than 1-gallon (approx. 3.8 L). If the concentration was >10% and was used for disinfecting, limit the sink disposal to less than 2-cups (approx. 500 mL) of bleach solution. Also flush the drain with adequate volumes of water.

This does not apply to “waste” bleach, e.g., unused or expired bleach. Any leftover, unused product or expired bleach not used as intended by the manufacturer, or used as a disinfectant, needs be collected by OSEH Hazardous Materials Management (HMM) for proper disposal. Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, and waste collection. Also refer to the OSEH Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==
If the employee is in need of emergency medical attention, call 911 immediately.

In general, flush affected eyes or skin with water for at least 15 minutes, then seek medical attention (see below).
* Eyes: In case of contact, immediately flush eyes with plenty of water for at least 15 minutes. Get medical aid.
* Skin: In case of contact, immediately flush skin with plenty of water. Remove contaminated clothing and shoes. Get medical aid. Wash clothing before reuse.
* Ingestion: Get medical aid immediately. Do not induce vomiting unless directed to do so by medical personnel. Never give anything by mouth to an unconscious person. If vomiting occurs naturally, have victim lean forward. Wash mouth with water, and then give plenty of milk or water to drink and obtain urgent medical attention.
* Inhalation: If inhaled, remove to fresh air. If not breathing, give artificial respiration. Get medical aid.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==Treatment Facilities==
* U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
* University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
* UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

==Spill Procedures==
* When a spill occurs, personal safety should always come first.
* Alert and clear everyone in the immediate area where the spill occurred.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., less than 1 Gallon or 3.5 Liters. A major/large chemical spill requires active assistance from emergency personnel.

For minor (small) spills
* Use proper personal protective equipment as indicated above.
* Absorb the spilled bleach using an absorbent, non-combustible material such as appropriate sorbent pads, sand, or vermiculite. Do not use combustible materials such as sawdust.
* Collect residue, place in container and contact OSEH-HMM (734) 763-4568 for proper disposal.

For major (large) spills
Report large chemical spills greater than 1 gallon or 3.5 liters in corridors or common areas, e.g., hallways, elevators, eating areas, rest rooms, offices, etc., to University of Michigan Division of Public Safety and Security (DPSS) by calling 911.

Additional Spill Response Steps:

===Minor Chemical Spill===
* Alert people in immediate area of spill.
* Open outside windows, if possible.
* Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.
* Avoid breathing vapors from spill.
* Confine spill to as small an area as possible.
* Do not wash spill down the drain.
* Use appropriate spill kits/sorbents to neutralize corrosives and/or absorb spill. Collect contaminated materials and residues and place in container. For powdered chemicals sweep carefully to avoid generation of dust or, if appropriate, use moist sorbent pads or wet the powder with a suitable solvent and then wipe with a dry cloth. Contact OSEH-HMM (734) 763-4568 for proper disposal.
* Clean spill area with water.

===Major Chemical Spill===
* Attend to injured or contaminated persons and remove them from exposure.
* Alert people in the laboratory to evacuate.
* Close doors to affected area.
* Post warnings to keep people from entering the area.
* Have person available that has knowledge of incident and laboratory to assist emergency personnel.

===Additional Spill Links===
* http://www.oseh.umich.edu/pdf/chemspil.pdf
* http://www.oseh.umich.edu/emer-chemical.shtml

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling bleach.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 16:42, 12 October 2016 (UTC)
* [[User:Ccousz|Ccousz]] ([[User talk:Ccousz|talk]])
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:17, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 18:47, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:45, 12 June 2017 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 14:52, 13 June 2017 (UTC)
*[[User:Nlambrec|Nlambrec]] ([[User talk:Nlambrec|talk]]) 21:08, 17 February 2019 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])
*[[User:elhabbal|elhabbal]] ([[User talk:elhabbal|talk]]) 7 October 2019
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☒ NO

Laboratory Director - Dave Bridges Revision Date - 2016-07-26

SOP - Phenol

2019-10-07T15:22:57Z

Elhabbal: /* Spill Procedure */

[[Category: SOP]]
[[Category: Lab Safety]]
'''Laboratory Director (LD) Approval is Required Prior to Performing this Procedure'''

=Description=
This standard operating procedure (SOP) outlines the handling and use of phenol. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling phenol.

Phenol is used as a reagent in chemical analysis. It causes burns and risk of serious damage to eyes. Phenol is toxic by inhalation, in contact with skin and if swallowed.

=Potential Hazards=
• Phenol is very hazardous in case of skin contact – it is corrosive, toxic, and can cause irritation. Initially, it can cause numbness or slight tingling, so employees may not be immediately aware of contact. If absorbed through the skin it can cause muscle weakness, tremors, loss of coordination, shock, sudden collapse, coma, convulsions, organ damage, and death. It may cause severe eye injury (including blindness) if it contacts the eyes, and is extremely toxic (fatal) by ingestion.

• Inhalation exposure is less likely – it does not evaporate easily at room temperature, but can be inhaled if heated and/or misted, or in the case of a large spill. If inhaled, phenol can cause upper respiratory irritation, lung damage, and CNS impairment.

• For more safety information, refer to Prudent Practice’s Laboratory Chemical Safety Summary for Phenol.

Occupational Exposure Limits (OELs):
• MIOSHA PEL and ACGIH TLV: 5 ppm, 8-hour TWA

=Engineering Controls=
An eyewash (preferably eyewash/drench hose combination unit) must be located in the immediate area. If medium-large quantities are used (>~100 ml of 100% phenol), a safety shower is also required. Contact OSEH at (734) 647-1143 for an assessment of the need for a safety shower if one is not available. Work with large open containers should be performed only in a chemical fume hood. Small amounts can be handled safely on the bench top, as long as skin contact is avoided.
�
=Work Practice Controls=
• Designate areas where phenol is stored or manipulated.

• It is highly recommended that labs using phenol (or reagents containing phenol – e.g., TRIzol) have polyethylene glycol 300 or 400 (PEG-300 or PEG-400) on hand in case of dermal exposure.
• Purchase in the smallest container that is practical for lab use.

• Purchase in a shatter-resistant container if available (such as PVC-coated glass).

• Keep containers closed as much as possible.

• Use in the smallest quantities and lowest concentration practicable for the experiment being performed.

• After work with phenol is complete, wipe down work area with soap and water solution.

=Personal Protective Equipment=
• Wear neoprene or doubled nitrile laboratory gloves, safety glasses, and lab coat. If gloves are splashed, remove them immediately and put on a new pair of gloves. If splashes may occur, wear a face shield with chemical splash goggles, and an impervious apron over the lab coat.

• If phenol and chloroform will be used together, please note that chloroform easily penetrates nitrile gloves and can degrade them, allowing phenol to contact the skin. Plan work to avoid glove contact and change gloves immediately if there is a splash. If extensive work with phenol and chloroform is done in the lab, consider the reusable ChemTek Viton/Butyl glove made for work with both of these chemicals (or they can be used for spill clean-up).

=Transportation and Storage=
• Transport phenol in secondary containment, preferably a polyethylene or other non-reactive acid/solvent bottle carrier.

• Keep container in cool, well-ventilated area.

• Keep container tightly closed and sealed until ready for use.

• Store in secondary containment away from moisture, strong oxidizers, strong caustics, plastics, rubber, nitric acid, water + heat, and chemically active metals, such as aluminum and magnesium powder, sodium, potassium, and lithium.

• Store below eye level.

• Avoid storing on the floor.

• Avoid ignition sources.

=Waste Disposal=
Handle and store hazardous waste following the guidelines above for work practice controls, transportation and storage. Because most spent, unused and expired chemicals/materials are considered hazardous wastes, they must be properly disposed of. Do not dispose of chemical wastes by dumping them down a sink, flushing in a toilet or discarding in regular trash containers, unless authorized by OSEH Hazardous Materials Management (HMM). Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.
�
=Exposures/Unintended contact=

If the employee is in need of emergency medical attention, call 911 immediately.

For an actual chemical exposure/injury:
• Remove contaminated clothing. Flush exposed eyes or skin with water for at least 15 minutes. Seek medical attention (see below).

• If a small area of skin has been exposed to phenol, swab polyethylene glycol 300 or 400 (PEG-300 or 400) onto the affected area immediately and repeatedly until the smell of phenol is no longer evident, then seek medical attention. If PEG-300 or 400 is not available, flush area with COPIOUS amounts of water (such as from a drench hose or safety shower) for at least 15 minutes, then seek medical attention. See the US Agency for Toxic Substances and Disease Registry (ATSDR), Medical Management Guidelines for Phenol.

• For larger areas of exposed skin or eye exposure, flush area with COPIOUS amounts of water for at least 15 minutes, and then seek immediate medical attention. Please note that using high-density water irrigation will reduce phenol uptake, but if lesser amounts of water are used it will merely dilute the phenol and increase the area of exposure.

• For situations with risk of inhalation exposure (including spills of powder outside of a chemical fume hood), remove all persons from the contaminated area.

• If an ambulance is needed, call the University of Michigan Division of Public Safety and Security (DPSS) at 911 to request assistance.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to Work Connections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the Work Connections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm

After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021

University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320

UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

�
=Spill Procedure=
• When a spill occurs, personal safety should always come first.

• Alert and clear everyone in the immediate area where the spill occurred.

• Employees in the area should be prepared to clean up minor spills, including most spills confined to the chemical fume hood. Wearing double nitrile gloves, splash goggles, face shield and lab coat (and impermeable apron, if available), use absorbent pads to absorb spilled material. After spill has been completely absorbed, wipe down contaminated area with soap and water solution. Lab personnel should take great care to avoid skin contact with phenol. If skin contact does occur, follow the instructions outlined for exposures/unintended contact in the section above.

• Contaminated PPE and clean-up materials should be placed in a clear plastic bag. Call OSEH-HMM (734) 763-4568 to arrange for waste pick-up.

• If a large spill occurs, leave the lab and call University of Michigan Division of Public Safety and Security (DPSS) at 911 to request assistance from OSEH. Do not take any action to cover the spill. Post a warning on the lab and do not allow others to enter. Have a person available that has knowledge of the incident and laboratory to assist emergency personnel.

Additional Spill Links:
• http://oseh.umich.edu/hazardous-waste/spill-response/

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

Training of Personnel
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling phenol.

Certification
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure.

*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 20:50, 10 November 2016 (UTC)
* [[User:Davebridges|Davebridges]] ([[User talk:Davebridges|talk]]) 21:08, 6 December 2016 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:47, 13 June 2017 (UTC)
*[[User:Cdgleason|Cdgleason]] ([[User talk:Cdgleason|talk]]) 03:55, 4 October 2018 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 21:43, 12 September 2019 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])
*[[User:elhabbal|elhabbal]] ([[User talk:elhabbal|talk]]) 7 October 2019
Laboratory Director Revision Date
�
Major Revisions (Tracking purposes only – Do not print as part of SOP)

Generating DMSO Stocks for Cell Culture

2018-06-01T18:34:57Z

Elhabbal:

==Materials==
*Cells in 10cm dishes, at 90-95% confluence.
*Cryopreservation Container (Nalgene 5100-0001)
*Cryopreservation Vials (Corning 430487)
*Sterile DMSO

==Protocol==
#Pick a low passage number of cells and grow 2-5 10cm dishes.
#At near confluence wash cells twice with PBS -/- and trypsinize normally.
#Collect all the cells in a 15 mL falcon tube. Add media up to 15 ml. (To avoid potentially over-trypsinizing, you can add media to each plate immediately after the cells detach, then add media up to the 15 ml mark on the falcon tube.)
#Centrifuge 5 min at 1500RPM to pellet cells.
#Aspirate media.
#Add media (1.8 mL per original plate, so if you started with 5 plates, the added media will be 5*1.8).
#Add sterile DMSO to a final concentration of 10% (0.2 mL per original plate, so if you started with 5 plates, the added DMSO will be 0.2*5).
#Gently resuspend cells and aliquot 1 mL of suspension into cryopreservation vials. (Resuspend with a pipette tip gently.)
#Label vials with name, date, cell type and passage (if known).
#Place container with vials at -80 for 1-3 days.
#Remove cells from container and place in liquid nitrogen storage.

[[Category: Cell Culture]]
[[Category: Storage]]

Generating DMSO Stocks for Cell Culture

2018-06-01T18:34:21Z

Elhabbal: clarified some calculations, removed the isopropanol step

==Materials==
*Cells in 10cm dishes, at 90-95% confluence.
*Cryopreservation Container (Nalgene 5100-0001)
*Cryopreservation Vials (Corning 430487)
*Sterile DMSO

==Protocol==
#Pick a low passage number of cells and grow 2-5 10cm dishes.
#At near confluence wash cells twice with PBS -/- and trypsinize normally.
#Collect all the cells in a 15 mL falcon tube. Add media up to 15 ml. (To avoid potentially over-trypsinizing, you can add media to each plate immediately after the cells detach, then add media up tot eh 15 ml mark on the falcon tube.)
#Centrifuge 5 min at 1500RPM to pellet cells.
#Aspirate media.
#Add media (1.8 mL per original plate, so if you started with 5 plates, the added media will be 5*1.8).
#Add sterile DMSO to a final concentration of 10% (0.2 mL per original plate, so if you started with 5 plates, the added DMSO will be 0.2*5).
#Gently resuspend cells and aliquot 1 mL of suspension into cryopreservation vials. (Resuspend with a pipette tip gently.)
#Label vials with name, date, cell type and passage (if known).
#Place container with vials at -80 for 1-3 days.
#Remove cells from container and place in liquid nitrogen storage.

[[Category: Cell Culture]]
[[Category: Storage]]

Preparation of RNA Samples from Mouse Tissues

2017-06-29T20:04:29Z

Elhabbal: Added where to dispose of the chloroform in case someone forgets.

==Safety Information==
* [[SOP-_Phenol|SOP - Phenol]]
* [[SOP_-_Chloroform|SOP - Chloroform]]

==Materials==
*PureLink RNA Mini Kit (Invitrogen cat#12183-018A)
*Mouse Tissue (50-100 mg, about a 3mm cube)
*TRIZol (Invitrogen cat# 12183-555)
*Chloroform (in solvent cabinet)
*Label tubes, for each sample need 3 eppies, at least one of which is 2 mL and one of which is a recovery tube from the PureLink kit.
*70% Ethanol make with RNAase free water and 100% Ethanol.

==Protocol==
#Add 1 mL TRIzol reagent to each 2 mL tube.
#Cut ~50-100 mg of tissue. If tissue is in RNAlater, cut at room temperature. If tissue is frozen cut on dry ice. Weigh into a fresh 2mL eppendorf tube and keep on ice.
#Using tissue grinder, and after adding the ball bearings to every vial, homogenize tissue for 3 min at 25Hz (make sure there are no remaining clumps).
#Incubate 5 minutes at room temperature.
#Add 200 uL Chloroform and shake vigourously by hand for 15s. Do not vortex.
#Transfer sample to a new 1.5 vial. The ball bearings are reusable after cleaning so do not discard them.
#Incubate at room temperature for 2-3 minutes.
#Centrifuge in a cold eppendorf centrifuge (4C) on maximum for 15 minutes. Sample will separate into a lower, red chloroform phase, an interphase and a colorless upper phase which contains the RNA.
#Add 400 uL of 70% ethanol to a fresh tube.
#Transfer 400 uL of the upper phase to the ethanol tube and mix by vortexing. The remaining chloroform in the previous vial should be disposed of in the phenol waste container under the hood.
#Remove 700 uL of mixture (invert briefly to mix before taking sample) and add to a spin column in a collection tube.
#Spin 15s on max. Discard flow through. Add remaining sample, respin and discard flow through.
#Add 700 uL Wash Buffer I to spin column.
#Spin 15s on max. Discard flow through the collection tube.
#Add 500 uL Wash Buffer II (make sure ethanol was added to the wash buffer) to the spin cartridge.
#Spin 15s on max. Discard the flow through and replace the collection tube.
#Repeat wash by adding 500 uL Wash Buffer II to the spin cartridge.
#Spin 15s on max. Discard the flow through and keep the same collection tube.
#Spin 1 min on max to dry the cartridge. Discard the collection tube and place into a clean recovery tube. Add 100 uL RNAase free water to the center of each tube. This can be adjusted to between 30 and 300 uL elution buffer if necessary.
#Incubate at room temperature for 1min.
#Spin 2 min at 12500 rpm to get purified RNA.
#Quantify the RNA using the nanodrop.

[[Category:qPCR]]
[[Category:Expression]]
[[Category:Transcription]]
[[Category:RNA]]
[[Category:Mouse Work]]
[[Category:Tissues]]

Preparation of RNA Samples from Mouse Tissues

2017-06-29T20:02:27Z

Elhabbal: Added ball bearing step, added step 6 to transfer to a new 1.5 vial.

==Safety Information==
* [[SOP-_Phenol|SOP - Phenol]]
* [[SOP_-_Chloroform|SOP - Chloroform]]

==Materials==
*PureLink RNA Mini Kit (Invitrogen cat#12183-018A)
*Mouse Tissue (50-100 mg, about a 3mm cube)
*TRIZol (Invitrogen cat# 12183-555)
*Chloroform (in solvent cabinet)
*Label tubes, for each sample need 3 eppies, at least one of which is 2 mL and one of which is a recovery tube from the PureLink kit.
*70% Ethanol make with RNAase free water and 100% Ethanol.

==Protocol==
#Add 1 mL TRIzol reagent to each 2 mL tube.
#Cut ~50-100 mg of tissue. If tissue is in RNAlater, cut at room temperature. If tissue is frozen cut on dry ice. Weigh into a fresh 2mL eppendorf tube and keep on ice.
#Using tissue grinder, and after adding the ball bearings to every vial, homogenize tissue for 3 min at 25Hz (make sure there are no remaining clumps).
#Incubate 5 minutes at room temperature.
#Add 200 uL Chloroform and shake vigourously by hand for 15s. Do not vortex.
#Transfer sample to a new 1.5 vial. The ball bearings are reusable after cleaning so do not discard them.
#Incubate at room temperature for 2-3 minutes.
#Centrifuge in a cold eppendorf centrifuge (4C) on maximum for 15 minutes. Sample will separate into a lower, red chloroform phase, an interphase and a colorless upper phase which contains the RNA.
#Add 400 uL of 70% ethanol to a fresh tube.
#Transfer 400 uL of the upper phase to the ethanol tube and mix by vortexing.
#Remove 700 uL of mixture (invert briefly to mix before taking sample) and add to a spin column in a collection tube.
#Spin 15s on max. Discard flow through. Add remaining sample, respin and discard flow through.
#Add 700 uL Wash Buffer I to spin column.
#Spin 15s on max. Discard flow through the collection tube.
#Add 500 uL Wash Buffer II (make sure ethanol was added to the wash buffer) to the spin cartridge.
#Spin 15s on max. Discard the flow through and replace the collection tube.
#Repeat wash by adding 500 uL Wash Buffer II to the spin cartridge.
#Spin 15s on max. Discard the flow through and keep the same collection tube.
#Spin 1 min on max to dry the cartridge. Discard the collection tube and place into a clean recovery tube. Add 100 uL RNAase free water to the center of each tube. This can be adjusted to between 30 and 300 uL elution buffer if necessary.
#Incubate at room temperature for 1min.
#Spin 2 min at 12500 rpm to get purified RNA.
#Quantify the RNA using the nanodrop.

[[Category:qPCR]]
[[Category:Expression]]
[[Category:Transcription]]
[[Category:RNA]]
[[Category:Mouse Work]]
[[Category:Tissues]]

Glycogen Determination from Tissues

2017-06-19T13:02:29Z

Elhabbal:

[[Category:Glycogen]]
[[Category:Metabolism]]
[[Category:Mouse Tissues]]
[[Category:Mouse Work]]

==Materials and Buffers==
* Screw Capped Vials
* 30% KOH, prepared fresh
* 1M Sodium Sulfate
* Ethanol
* 50 mM Sodium Acetate, pH 4.8
* Amyloglucosidase 0.3 mg/mL in 50 mM Sodium Acetate. Stored in -80. (Sigma A7420-5MG)
* Glucose quantification kit (Wako Autokit Glucose Buffer Solution cat # 439-9091; for protocol see http://www.wakodiagnostics.com/pi/pi_autokit_glucose.pdf)
* Glucose standard solution (200 or 500 mg/dL; Wako)

==Protocol==
# Weigh out 30-90 mg tissue into a '''screw cap vial''' and record weights. Screw cap vials are really important or else the lids will pop off.
# Turn on the heating block and set it to 95C (this can take up to 15 minutes to reach the desired temperature).
# Add 300 uL 30% KOH and place on 95C heat block for 30 min with occasional mixing by gently tapping the vials to ensue the tissue dissolves completely. Make sure all the sample is initially immersed in KOH.
# Cool and then add 100 uL Sodium Sulfate and 800 uL Ethanol.
# Boil for 5 min.
# Turn on the incubator and set it to 37C.
# Centrifuge at 13 000 RPM for 5 min.
# Aspirate the solution leaving the pellet at the bottom of the vial.
# Resuspend pellet in 200 uL water while making sure that all the glycogen dissolves in water, then add 400 uL ethanol.
# Boil 5 min, spin 5 min and Repeat wash steps twice more (Wash Steps: Aspirate -> Resuspend with H20 and EtOH -> Boil -> Centrifuge -> Aspirate).
# Dry pellet on the bench by leaving the vial cap open.
# Prepare amyloglucosidase solution by diluting the AG stock 100X into 50 mM Sodium Acetate, pH 4.8. Prepare enough for 200 uL per tube plus some extras
# Resuspend pellets in 200uL amyloglucosidase solution and incubate at 37C for 3h-O/N
# Quantify glucose using kit:
## Add 100 uL of Glucose buffer solution to each well, including wells for standard curve and blank
## Add 1-5 ul of glucose standard 200mg/dL diluted 1:5
## Add 10 uL digested glycogen for fasted samples, and use a 10X dilution for refed tissues (using either 5 or 2ul from the diluted sample)
## Mix and incubate at 37C for 5 min
## Measure absorbance at 505 nm
## Calculate glycogen levels as umoles glucose released per g tissue (should be ~10)

==Calculations==
# Use this Sweave template: https://raw.github.com/davebridges/biomolecule-scripts/master/R/Sweave/glycogen-analysis-tissue.Rnw
Reference:

PMID 15282316

Glycogen Determination from Tissues

2017-06-14T20:58:28Z

Elhabbal: Added step 2,clarified how to mix sample, added clarification to step 3 as to ensure all sample is immersed in KOH, added step 6, clarified the washing steps in order, clarified how to dry pellet

[[Category:Glycogen]]
[[Category:Metabolism]]
[[Category:Mouse Tissues]]
[[Category:Mouse Work]]

==Materials and Buffers==
* Screw Capped Vials
* 30% KOH, prepared fresh
* 1M Sodium Sulfate
* Ethanol
* 50 mM Sodium Acetate, pH 4.8
* Amyloglucosidase 0.3 mg/mL in 50 mM Sodium Acetate. Stored in -80. (Sigma A7420-5MG)
* Glucose quantification kit (Wako Autokit Glucose Buffer Solution cat # 439-9091; for protocol see http://www.wakodiagnostics.com/pi/pi_autokit_glucose.pdf)
* Glucose standard solution (200 or 500 mg/dL; Wako)

==Protocol==
# Weight out 30-90 mg tissue into a '''screw cap vial''' and record weights. Screw cap vials are really important or else the lids will pop off.
# Turn on the heating block and set it to 95C (this can take up to 15 minutes to reach the desired temperature).
# Add 300 uL 30% KOH and place on 95C heat block for 30 min with occasional mixing by gently tapping the vials to ensue the tissue dissolves completely. Make sure all the sample is initially immersed in KOH.
# Cool and then add 100 uL Sodium Sulfate and 800 uL Ethanol.
# Boil for 5 min.
# Turn on the incubator and set it to 37C.
# Centrifuge at 13 000 RPM for 5 min.
# Aspirate the solution leaving the pellet at the bottom of the vial.
# Resuspend pellet in 200 uL water while making sure that all the glycogen dissolves in water, then add 400 uL ethanol.
# Boil 5 min, spin 5 min and Repeat wash steps twice more (Wash Steps: Aspirate -> Resuspend with H20 and EtOH -> Boil -> Centrifuge -> Aspirate).
# Dry pellet on the bench by leaving the vial cap open.
# Prepare amyloglucosidase solution by diluting the AG stock 100X into 50 mM Sodium Acetate, pH 4.8. Prepare enough for 200 uL per tube plus some extras
# Resuspend pellets in 200uL amyloglucosidase solution and incubate at 37C for 3h-O/N
# Quantify glucose using kit:
## Add 700 uL Glucose Buffer Solution with Color Reagent to a plastic cuvette. (for microplate add 100ul)
## Add 1-5 uL glucose standard (500mg/dL) for standard curve (for microplate add 1-5 ul of glucose standard 200mg/dL diluted 1:5)
## Add 10 uL digested glycogen (for mice fasted more than 6 hours. for fed/short fast need less)
## Mix and incubate at 37C for 5 min
## Measure absorbance at 505 nm
## Calculate glycogen levels as umoles glucose released per g tissue (should be ~10)

==Calculations==
# Use this Sweave template: https://raw.github.com/davebridges/biomolecule-scripts/master/R/Sweave/glycogen-analysis-tissue.Rnw
Reference:

PMID 15282316

SOP - Phenol

2017-06-13T14:46:25Z

Elhabbal:

[[Category: SOP]]
[[Category: Lab Safety]]
'''Laboratory Director (LD) Approval is Required Prior to Performing this Procedure'''

=Description=
This standard operating procedure (SOP) outlines the handling and use of phenol. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling phenol.

Phenol is used as a reagent in chemical analysis. It causes burns and risk of serious damage to eyes. Phenol is toxic by inhalation, in contact with skin and if swallowed.

=Potential Hazards=
• Phenol is very hazardous in case of skin contact – it is corrosive, toxic, and can cause irritation. Initially, it can cause numbness or slight tingling, so employees may not be immediately aware of contact. If absorbed through the skin it can cause muscle weakness, tremors, loss of coordination, shock, sudden collapse, coma, convulsions, organ damage, and death. It may cause severe eye injury (including blindness) if it contacts the eyes, and is extremely toxic (fatal) by ingestion.

• Inhalation exposure is less likely – it does not evaporate easily at room temperature, but can be inhaled if heated and/or misted, or in the case of a large spill. If inhaled, phenol can cause upper respiratory irritation, lung damage, and CNS impairment.

• For more safety information, refer to Prudent Practice’s Laboratory Chemical Safety Summary for Phenol.

Occupational Exposure Limits (OELs):
• MIOSHA PEL and ACGIH TLV: 5 ppm, 8-hour TWA

=Engineering Controls=
An eyewash (preferably eyewash/drench hose combination unit) must be located in the immediate area. If medium-large quantities are used (>~100 ml of 100% phenol), a safety shower is also required. Contact OSEH at (734) 647-1143 for an assessment of the need for a safety shower if one is not available. Work with large open containers should be performed only in a chemical fume hood. Small amounts can be handled safely on the bench top, as long as skin contact is avoided.
�
=Work Practice Controls=
• Designate areas where phenol is stored or manipulated.

• It is highly recommended that labs using phenol (or reagents containing phenol – e.g., TRIzol) have polyethylene glycol 300 or 400 (PEG-300 or PEG-400) on hand in case of dermal exposure.
• Purchase in the smallest container that is practical for lab use.

• Purchase in a shatter-resistant container if available (such as PVC-coated glass).

• Keep containers closed as much as possible.

• Use in the smallest quantities and lowest concentration practicable for the experiment being performed.

• After work with phenol is complete, wipe down work area with soap and water solution.

=Personal Protective Equipment=
• Wear neoprene or doubled nitrile laboratory gloves, safety glasses, and lab coat. If gloves are splashed, remove them immediately and put on a new pair of gloves. If splashes may occur, wear a face shield with chemical splash goggles, and an impervious apron over the lab coat.

• If phenol and chloroform will be used together, please note that chloroform easily penetrates nitrile gloves and can degrade them, allowing phenol to contact the skin. Plan work to avoid glove contact and change gloves immediately if there is a splash. If extensive work with phenol and chloroform is done in the lab, consider the reusable ChemTek Viton/Butyl glove made for work with both of these chemicals (or they can be used for spill clean-up).

=Transportation and Storage=
• Transport phenol in secondary containment, preferably a polyethylene or other non-reactive acid/solvent bottle carrier.

• Keep container in cool, well-ventilated area.

• Keep container tightly closed and sealed until ready for use.

• Store in secondary containment away from moisture, strong oxidizers, strong caustics, plastics, rubber, nitric acid, water + heat, and chemically active metals, such as aluminum and magnesium powder, sodium, potassium, and lithium.

• Store below eye level.

• Avoid storing on the floor.

• Avoid ignition sources.

=Waste Disposal=
Handle and store hazardous waste following the guidelines above for work practice controls, transportation and storage. Because most spent, unused and expired chemicals/materials are considered hazardous wastes, they must be properly disposed of. Do not dispose of chemical wastes by dumping them down a sink, flushing in a toilet or discarding in regular trash containers, unless authorized by OSEH Hazardous Materials Management (HMM). Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.
�
=Exposures/Unintended contact=

If the employee is in need of emergency medical attention, call 911 immediately.

For an actual chemical exposure/injury:
• Remove contaminated clothing. Flush exposed eyes or skin with water for at least 15 minutes. Seek medical attention (see below).

• If a small area of skin has been exposed to phenol, swab polyethylene glycol 300 or 400 (PEG-300 or 400) onto the affected area immediately and repeatedly until the smell of phenol is no longer evident, then seek medical attention. If PEG-300 or 400 is not available, flush area with COPIOUS amounts of water (such as from a drench hose or safety shower) for at least 15 minutes, then seek medical attention. See the US Agency for Toxic Substances and Disease Registry (ATSDR), Medical Management Guidelines for Phenol.

• For larger areas of exposed skin or eye exposure, flush area with COPIOUS amounts of water for at least 15 minutes, and then seek immediate medical attention. Please note that using high-density water irrigation will reduce phenol uptake, but if lesser amounts of water are used it will merely dilute the phenol and increase the area of exposure.

• For situations with risk of inhalation exposure (including spills of powder outside of a chemical fume hood), remove all persons from the contaminated area.

• If an ambulance is needed, call the University of Michigan Division of Public Safety and Security (DPSS) at 911 to request assistance.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to Work Connections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the Work Connections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm

After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021

University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320

UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

�
=Spill Procedure=
• When a spill occurs, personal safety should always come first.

• Alert and clear everyone in the immediate area where the spill occurred.

• Employees in the area should be prepared to clean up minor spills, including most spills confined to the chemical fume hood. Wearing double nitrile gloves, splash goggles, face shield and lab coat (and impermeable apron, if available), use absorbent pads to absorb spilled material. After spill has been completely absorbed, wipe down contaminated area with soap and water solution. Lab personnel should take great care to avoid skin contact with phenol. If skin contact does occur, follow the instructions outlined for exposures/unintended contact in the section above.

• Contaminated PPE and clean-up materials should be placed in a clear plastic bag. Call OSEH-HMM (734) 763-4568 to arrange for waste pick-up.

• If a large spill occurs, leave the lab and call University of Michigan Division of Public Safety and Security (DPSS) at 911 to request assistance from OSEH. Do not take any action to cover the spill. Post a warning on the lab and do not allow others to enter. Have a person available that has knowledge of the incident and laboratory to assist emergency personnel.

Additional Spill Links:
• http://oseh.umich.edu/hazardous-waste/spill-response/

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

Training of Personnel
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling phenol.

Certification
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure.

*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 20:50, 10 November 2016 (UTC)
* [[User:Davebridges|Davebridges]] ([[User talk:Davebridges|talk]]) 21:08, 6 December 2016 (UTC)

Laboratory Director Revision Date
�
Major Revisions (Tracking purposes only – Do not print as part of SOP)

SOP - Vacuum Pumps

2017-06-12T22:25:19Z

Elhabbal: /* Training of Personnel */

[[Category: SOP]]
[[Category: Lab Safety]]

==Description==
This standard operating procedure outlines the handling and use of vacuum pumps. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when working with vacuum pumps.

Vacuum pumps are commonly used in a variety of experimental setups to remove air and other vapors from a vessel or system. Applications that use vacuum pumps include rotary evaporators, vacuum ovens, drying manifolds, freeze-dryers, aspirators, desiccators, and filtration equipment. Many vacuum pumps use oil which can be easily contaminated and result in pump damage and hazards.

==Potential Hazards==
Vacuum pumps can pose chemical, mechanical, electrical, and fire hazards. Chemical exposure can occur from improper installing, trapping, and exhausting or from off gassing of contaminated pump oil. Mechanical hazards can occur from the moving parts of the vacuum pump. Fire can result if the pump malfunctions or overheats and ignites nearby combustible materials. Electrocution can occur from faulty or defective switches and wiring.

==Engineering Controls==
Vacuum pumps used to evacuate systems containing toxic, volatile, or corrosive substances must be vented to the building exhaust ventilation system. Whenever possible locate pump in vacuum pump cabinet or a ventilated cabinet. Ensure that pumps have belt guards. Traps such as a cold trap should be used in line with high vapor loads to minimize the amount of volatile chemicals being evacuated and reaching the pump oil. Ensure the cold trap is appropriate for situation and follows all manufacturer and safety guidelines.

==Work Practice Controls==
===General:===
* Place pump on spill tray to contain oil spills if the equipment fails.
* Shield any glassware under vacuum.
===Physical:===
* Ensure all vacuum pump belt systems are guarded.
* Electrical cords and switches must be free from defects.
* Avoid placing pumps in an unventilated and enclosed receptacle.
* Do not operate pumps near combustible materials.
* Use correct vacuum tubing and replace old tubing.
===Chemical:===
* Use engineering controls to avoid exposure.
* Ensure pump oil is compatible with vapors that will pass through the pump (i.e. avoid hydrocarbon pump oil and oxidizing gases/vapors).
* Close valve between vessel and pump before turning off pump to avoid introducing oil into system.
* Ensure gases or vapors will not damage the pump.
* If cold traps are used, check for blockage.
* Check oil levels and change oil when necessary. Change oil when it begins to turn a dark brown color.
* An oil mist separator can be used to prevent oil loss.

==Personal Protective Equipment (PPE)==
Wear safety glasses, lab coat, long pants, close-toed shoes and gloves when performing all vacuum operations.

==Transportation and Storage==
Vacuum pumps must be stored on spill trays to prevent oil spills. Vacuum pump areas may be insulated for noise, if necessary.

==Waste Disposal==
All used vacuum pump oils must be dispose of through OSEH.

Do not dispose of chemical wastes by dumping them down a sink, flushing in a toilet or discarding in regular trash containers, unless authorized by OSEH Hazardous Materials Management (HMM). Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===
===U-M Occupational Health Services -- Campus Employees===
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
===University Health Services -- University students (non-life threatening conditions)===
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
===UMHS Emergency Department -- after clinic hours or on weekends===
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for additional accident and injury reporting information.

==Spill Procedure==
A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., less than 1 Gallon or 3.5 Liters. A major/large chemical spill requires active assistance from emergency personnel.

For small oil spills, treat the oil as a hazardous chemical spill. Use appropriate absorbent and dispose of as hazardous waste. For large oil spills contact OSEH-HMM at (734) 763-4568.

* When a spill occurs, personal safety should always come first.
* Alert and clear everyone in the immediate area where the spill occurred.

Additional Spill Response Steps:

===MINOR CHEMICAL SPILL===
* Alert people in immediate area of spill.
* If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
* Open outside windows, if possible.
* Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.
* Avoid breathing vapors from spill.
* Confine spill to as small an area as possible.
* Do not wash spill down the drain.
* Use appropriate spill kits/sorbents to absorb spill. Collect contaminated materials and residues and place in container. Contact OSEH-HMM (734) 763-4568 for proper disposal.
* Clean spill area with water.

===MAJOR CHEMICAL SPILL===
* Attend to injured or contaminated persons and remove them from exposure.
* Alert people in the laboratory to evacuate.
* If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
* Call University of Michigan Division of Public Safety and Security (DPSS) at 911 immediately for assistance.
* Close doors to affected area.
* Post warnings to keep people from entering the area.
* Have person available that has knowledge of incident and laboratory to assist emergency personnel.

Additional Spill Links:
* www.oseh.umich.edu/pdf/chemspil.pdf
* http://www.oseh.umich.edu/emer-chemical.shtml.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when working with vacuum pumps.

Certification
I have read and understand the above SOP. I agree to contact my Supervisor or Lab Manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO

*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 16:18, 31 October 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:27, 10 November 2016 (UTC)
*[[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:56, 7 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:05, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:25, 12 June 2017 (UTC)

Laboratory Director - Dave Bridges Revision Date - 2016-10-19

SOP - Tamoxifen

2017-06-12T22:23:52Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety]]
'''Laboratory Director (LD) Approval is Required Prior to Performing this Procedure'''

=Description=
This standard operating procedure (SOP) outlines the handling and use of tamoxifen. Review this document and supply the information required in order to make it specific to your facility. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling tamoxifen.

Tamoxifen is classified as a hazardous drug according to the National Institute for Occupational Safety and Health (NIOSH). Tamoxifen is a triphenyl ethylene derivative with marked anti-estrogen properties, which binds to estrogen receptors and, as a result, has proven useful in the treatment of breast cancer (antineoplastic). Tamoxifen citrate has been used to stimulate ovulation in infertility.

Synonyms: Nolvadex, Istubal, Valodex

=Potential Hazards=
• May cause cancer.

• May impair fertility and cause harm to the unborn child or breastfed babies.

• Accidental ingestion of the material may be harmful.

• May cause eye irritation.

• This product may be harmful if inhaled and cause respiratory tract irritation. Persons with impaired respiratory function, airway diseases, and conditions such as emphysema or chronic bronchitis, may incur further disability if excessive concentrations of particulate are inhaled.

• The material is not thought to be a skin irritant (as classified using animal models). But may be harmful if absorbed through the skin and may cause skin irritation; systemic effects may result following absorption. Entry into the blood-stream through, for example, cuts, abrasions or lesions, may produce systemic injury with harmful effects. Examine the skin prior to the use of the material and ensure that any external damage is suitably protected.

=Engineering Controls=
• Use a biological safety cabinet or chemical fume hood during reconstitution and dilution of this material.

• Syringes used for tamoxifen injection must be safety engineered (self-sheathing syringes, luer-lock syringes, etc.)

• Use with animals typically requires ABSL2 containment housing.

• Animals should be appropriately restrained and/or sedated prior to administering injections and other dosing methods.

• Laboratories and other spaces where handling of tamoxifen occurs must be equipped with an eyewash station.

=Work Practice Controls=
• Avoid inhalation.

• Good hygiene practice requires that exposure be kept to a minimum and that suitable PPE and control measures be used in an occupational setting.

• Decontamination should consist of surface cleaning with water and detergent followed by thorough rinsing. The use of detergent is recommended because there is no single accepted method of chemical deactivation for all agents involved.

• A plastic backed absorbent pad should be placed under the work area during the process. This should be changed at the end of each procedure or when a spill occurs.

=Personal Protective Equipment (PPE)=
Appropriate lab attire including long pants and closed toe shoes must be worn. Personal protective equipment includes a lab coat or disposable coveralls, eye protection and two pairs of disposable nitrile gloves. Gloves should be changed frequently and should be pulled up over sleeves if possible to reduce the likelihood of any exposed skin. Wash hands and arms immediately after working with the substance.

Note: Respirators are masks designed to protect the wearer from specific airborne hazards and are different from surgical masks, which protect the wearer only from splashes and are primarily intended to protect others from infectious aerosols exhaled by the wearer. Respirator use requires employee participation in the Respiratory Protection Program, which involves medical clearance and annual fit testing and training. Please be clear about use of surgical masks versus respirators. (Do NOT use the vague term “masks”.)

=Transportation and Storage=
• Keep containers tightly closed and in dry, well-ventilated area.

• Keep away from light.

• Avoid strong oxidizing agents.

• Tamoxifen is regulated by DOT as a Hazardous Material as well as IATA for Transport of Dangerous Goods.

=Waste Disposal=
All hazardous chemical waste should be placed in an appropriate container for disposal (pail or bottle). The container should be closed except when actively adding waste. The container must be located in the area where hazardous drugs are being used.

All items contaminated by hazardous drugs, including gloves, syringes, vials needles, animal bedding and solution containers should be disposed according to OSEH guidelines.

• Sharps – place needles, syringes with needles attached and other breakable items into appropriately labeled sharps containers.

• Empty stock vials, reagent bottles, etc. – triple rinse with copious amounts of water. Deface label with black magic marker or scraper. Place in a cardboard box for disposal.

Because most spent, unused and expired chemicals/materials are considered hazardous wastes, they must be properly disposed of. Do not dispose of chemical wastes by dumping them down a sink, flushing in a toilet or discarding in regular trash containers, unless authorized by OSEH Hazardous Materials Management (HMM). Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

=Exposures/Unintended Contact=

If the employee is in need of emergency medical attention, call 911 immediately.

==SWALLOWED==
• Rinse mouth with water.

• Consult a physician.

==EYE==
If this product comes in contact with the eyes:
• Wash out immediately with fresh running water for a minimum of 15 minutes.

• Ensure complete irrigation of the eye by keeping eyelids apart and away from eye and moving the eyelids by occasionally lifting the upper and lower lids.

• If pain persists or recurs seek medical attention.

• Removal of contact lenses after an eye injury should only be undertaken by skilled personnel.

==SKIN==
If skin contact occurs:
• Immediately remove all contaminated clothing, including footwear.

• Flush skin and hair with running water (and soap if available) for a minimum of 15 minutes.

• Seek medical attention in event of irritation.

==INHALED==
• Move person from contaminated area into fresh air.

• Move person into fresh air.

• Seek medical attention.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to Work Connections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the Work Connections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm

After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021

University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320

UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

=Spill Procedure=
• When a spill occurs, personal safety should always come first.

• Alert and clear everyone in the immediate area where the spill occurred.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., (less than 1 Gallon or 3.5 Liters). A major/large chemical spill requires active assistance from emergency personnel.

Spill Response Steps:

===MINOR CHEMICAL SPILL===
• Alert people in immediate area of spill.

• Open outside windows, if possible.

• Use proper personal protective equipment (PPE) as indicated above.

• Avoid breathing vapors from spill.

• Confine spill to as small an area as possible.

• Do not wash spill down the drain.

• Use appropriate spill kits/sorbents to absorb spill. Collect contaminated materials and residues and place in container. For powdered chemicals sweep carefully to avoid generation of dust or, if appropriate, use moist sorbent pads or wet the powder with a suitable solvent and then wipe with a dry cloth. Contact OSEH-HMM (734) 763-4568 for proper disposal.

• Clean spill area with water.

===MAJOR CHEMICAL SPILL===
Report large chemical spills (greater than 1 Gallon or 3.5 Liters) in corridors or common areas, e.g., hallways, elevators, eating areas, rest rooms, offices, etc., to University of Michigan Division of Public Safety and Security (DPSS) at 911.
• Attend to injured or contaminated persons and remove them from exposure.

• Alert people in the laboratory to evacuate.

• Call University of Michigan Division of Public Safety and Security (DPSS) at 911 immediately for assistance.

• Close doors to affected area.

• Post warnings to keep people from entering the area.

• Have person available that has knowledge of incident and laboratory to assist emergency personnel.

Additional Spill Links:
• http://oseh.umich.edu/hazardous-waste/spill-response/

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

=Training of Personnel=
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling tamoxifen.

=Certification=
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure.

*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 20:56, 10 November 2016 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:23, 12 June 2017 (UTC)

Laboratory Director Revision Date
�
Major Revisions (Tracking purposes only – Do not print as part

SOP - Sensitizers

2017-06-12T22:22:29Z

Elhabbal: /* Certification */

[[Category: SOP]]
[[Category: Lab Safety]]

==Description==
This standard operating procedure outlines the handling and use of sensitizers. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling sensitizers.

==Potential Hazards==
A sensitizer (allergen) is a substance that causes exposed people to develop an allergic reaction in normal tissue after repeated contact with the substance. The reaction might be as mild as a rash (contact dermatitis) or as serious as anaphylactic shock. Sensitizers may also be corrosive or carcinogenic.

Examples of compounds that may cause sensitization in some individuals are diazomethane, various isocyanates, formaldehyde, latex, and benzylic and allylic halides.

==Engineering Controls==
Use a properly functioning lab fume hood when handling sensitizers that can be inhaled (via mist/fume/gas/vapor).
If the process does not permit the handing of such materials in a fume hood, contact OSEH at (734) 647-1143 for review of the adequacy of ventilation measures.

==Work Practice Controls==
For sensitizers that are also considered a particularly hazardous substances, a designated area shall be established per the other applicable SOP.
*The use of latex gloves may sensitize you to latex. If you feel that you develop a reaction to latex, please inform the PI and nitrile gloves will be provided for you.
*The use of masks while in the animal rooms may help prevent sensitizing to animal dander. These are provided in each animal room and are highly recommended for use each time interacting with the animals.

==Personal Protective Equipment (PPE)==
At minimum, safety glasses, lab coat, long pants, and closed toed shoes are to be worn when entering laboratories having hazardous chemicals.

Additionally, when handling hazardous chemicals or contacting potentially contaminated surfaces, protective gloves are to be worn. For proper selection of glove material, review the SDS or refer to the glove compatibility charts available through the OSEH website.

For hazardous chemicals that are toxic via skin contact/ absorption, additional protective clothing (i.e., face shield, apron, oversleeves) is appropriate where chemical contact with body/skin is foreseeable.

Goggles (not safety glasses) are appropriate for processes where splash or spray is foreseeable.

==Transportation and Storage==
Follow any substance-specific storage guidance provided in the SDS documentation.
�
==Waste Disposal==
Many sensitizers intended for disposal may likely be considered hazardous wastes. For general guidance regarding waste disposal, refer to the OSEH Hazardous Materials Management (HMM) chemical waste information available online. For specific guidance, contact OSEH-HMM at (734) 763-4568.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.

Contact Occupational Health Services (734) 764-8021 or the UMHS Emergency Room (734) 936-6666 for advice. Be prepared to provide them with a copy of the SDS so that medical personnel are aware of the material(s) involved and take precautions to protect themselves.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==TREATMENT FACILITIES:==
*U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
*University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
*UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for additional accident and injury reporting information.

==Spill Procedure==
*When a spill occurs, personal safety should always come first.
*Alert and clear everyone in the immediate area where the spill occurred.

For a small spill, inside the fume hood, use proper personal protective equipment as indicated above. Absorb the material with an absorbent, non-combustible material such as sorbent pads, sand or vermiculite. Collect residue, place in container and contact OSEH-HMM (734) 763-4568 for proper disposal.

==MAJOR CHEMICAL SPILL==
*Attend to injured or contaminated persons and remove them from exposure.
*Alert people in the laboratory to evacuate.
*If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
*Call University of Michigan Division of Public Safety and Security (DPSS) at 911 immediately for assistance.
*Close doors to affected area.
*Post warnings to keep people from entering the area.
*Have person available that has knowledge of incident and laboratory to assist emergency personnel.

==Additional Spill Links:==
*www.oseh.umich.edu/pdf/chemspil.pdf
*http://www.oseh.umich.edu/emer-chemical.shtml.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling sensitizers.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 16:13, 19 October 2016 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 16:55, 7 November 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:27, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:53, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:22, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO
Laboratory Director - Dave Bridges Revision Date - 2016-10-19

Spill Response Procedures for Infectious agents and Recombinant DNA

2017-06-12T22:21:00Z

Elhabbal: /* Certification */

Spills and accidents should be immediately reported to the Principal Investigator and the Department of Occupational Safety and Environmental Health (OSEH) (647-1143).

Large Spills Outside of the Biosafety Cabinet
OSEH will respond to large spills (greater than 200 ml) of BSL-2 (or above) containment level material. Contact OSEH (647-1143) for help with large spill cleanup.
Small Spills Outside of the Biosafety Cabinet
# Don double gloves and eye protection if not already wearing them.
# Cover spilled material with an absorbent paper towel or Kimwipe. Once the absorbent material is in place over the spill, wet the material with a 10% solution of bleach (1:10 dilution of Clorox containing sodium hypochlorite) or other appropriate disinfectant.
# Let stand 15-20 minutes, wipe up and wash surface with appropriate disinfectant.
# Wipe down all equipment and surfaces which may have been splashed.
Spill in a Biosafety Cabinet
Note: Leave the cabinet turned on.
# Don double gloves, a lab coat, and eye protection if not already wearing them.
# Cover spilled material with an absorbent paper towel or Kimwipe. Once the absorbent material is in place, wet material with 10% solution of bleach (1:10 dilution of Clorox containing sodium hypochlorite) or other appropriate disinfectant. Let stand 15-20 minutes, wipe up and wash surface with appropriate disinfectant.
# If personnel are contaminated, remove potentially contaminated garments at the BSC and decontaminate garments by saturation with 70% ethanol or place in autoclave bag for autoclaving. Wash hands and other potentially exposed skin surfaces thoroughly with soap and water. Don fresh PPE, return to worksite, and spray walls, liners, and equipment with an appropriate disinfectant.
# Spray or wipe container walls, other work surfaces and equipment with the appropriate disinfectant
# If necessary, flood the work surface, drain pan and catch basin below the work surface with disinfectant. Allow at least 15-20 minutes contact time.
# Soak up the disinfectant and drain the catch basin into a container. Lift the front exhaust grille and tray and wipe all surfaces. Ensure that no foreign materials are blown into the area below the grille.
# If a 10% bleach solution is used on metal surfaces, rinse with water or 70% ethanol after decontamination is complete.
# If the spill overflows into the interior of the cabinet, more extensive decontamination of the cabinet may be necessary. Contact OSEH (763-6973) for decontamination of the cabinet.
Spill in the Laboratory
# If an infectious aerosol may exist outside of a biosafety cabinet, leave the room immediately, lock the door, post a warning sign and inform your supervisor. If clothing is contaminated, remove and turn the exposed side of fabric in on itself and place in autoclave bag or biohazard container. Wait at least 30 minutes before reentering the lab to allow dissipation of aerosol created by the spill. During this time, review clean-up procedures, assemble decontamination materials, PPE and contact OSEH (647-1143).
# Don fresh gloves, a lab coat or tyvek suit, and eye protection.
# Carefully lay disinfectant-soaked towels over the spill and pour more around the spill. Use more concentrated disinfectant if the volume of material will significantly dilute the disinfectant.
# Allow 15-20 minutes contact time.
# Use forceps to place sharp objects into a sharps container. Using a dustpan and dustbroom, tongs, etc., transfer all contaminated materials (paper towels, gloves, labware, etc.) biohazard waste containers and contact OSEH HazMat (763-4568) for removal.
# Wipe surrounding surfaces with disinfectant to cover all splash areas. Wipe flat surfaces to remove any material that may have splashed out and settled on those surfaces.
# Place all contaminated materials, including protective clothing, into an autoclave bag or biohazard waste container.
# Wash hands with soap and warm water.
# If a personnel exposure occurred, complete an Illness or Injury Report form (https://www.workconnections.umich.edu/illnessorinjury.php) summit to Work~Connections and report incident to OSEH (763-6973). See additional procedures noted below.
Skin, Mucous Membrane, or Injury Exposure
# If there is an agent-specific protocol for exposures, follow that (e.g., HIV, Herpes B).
# In the case of skin contact or injury with a contaminated instrument:
## Thoroughly wash area with soap and water. Do not squeeze the wound to induce bleeding.
## Avoid use of abrasive chemical soaps or disinfectant washes as they can cause skin abrasions and a possible additional route of entry for the agent.
## For mucous membranes (e.g., eyes, mouth), flush for a minimum of 15 minutes.
# Notify UM-Occupational Health Services (OHS) for evaluation of exposure following:
## Contact with mucous membranes;
## Contact with non-intact skin;
## Percutaneous exposure; or
## Any type of exposure that involves concentrated virus.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Davebridges|Davebridges]] ([[User talk:Davebridges|talk]]) 14:44, 13 September 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:28, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:58, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:21, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO

Laboratory Director - Dave Bridges
Revision Date - 2016-09-13

[[ Category:SOP ]]

SOP - Biosafety Cabinets

2017-06-12T22:18:04Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety]]
=Description=
This standard operating procedure outlines the use of biosafety cabinets. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate administrative controls and personal protective equipment using biosafety cabinets.

Biological Safety Cabinets, (BSCs), also known as tissue culture hoods, are designed to provide personnel, environmental and product protection when appropriate practices and procedures are followed. Class II BSCs rely on directional movement of air to provide containment. Airflow is drawn into the front grille of the cabinet, providing personnel protection. The most commonly used BSC is a Class II A2. This type of cabinet is not suitable for volatile solvents.

BSC installation, required annual certification, decontamination and maintenance must be done by certified (accredited by the National Sanitation Foundation) professionals or by a certified OSEH technician. Maintenance and certification will be done in accordance with OSEH Engineering Controls Standard of Care #1-6.

=Work Practice Controls=
==Personal Protective Equipment:==
• Appropriate personal protective equipment (PPE) must be worn. Lab coats must be buttoned. Gloves should be pulled over the wrists of lab coat, not worn inside coat. Additional PPE to be used as recommended.

==Preparing BSC for work:==
• Confirm BSC annual (within 12 months) certification is current; information found on sticker on front of BSC.

• Operate cabinet blowers at least 3-5 minutes before beginning work to allow the BSC to “purge” particulates.

• Use 70% ethanol to clean work surface of BSC and to disinfect any glass, etc. that is being used; amount of alcohol in BSC must be for only one day’s work.

==Working in the BSC:==
¥ When working in the cabinet, move arms in and out slowly, perpendicular to the face opening to reduce disruption of air curtain.

¥ Perform all operations at least 4 inches from the front grille on the work surface.

¥ For BSC clean-up, apply 70% ethanol using wipes vs. spray bottles to minimize solvent vapor concentrations being re-circulated in the hood. Cabinet sash to remain open to allow for alcohol evaporation; sash can be lowered after sufficient time. The recommended minimum time for sash opening is 10 minutes.

¥ Do not bring potentially contaminated materials out of the cabinet until they have been surface decontaminated.

¥ Disposable underpads can be placed on the work surface but must not cover the front or rear grille openings. The use of toweling facilitates routine cleanup and reduces splatter and aerosol generation during an overt spill.

¥ Place all material as far back in the cabinet as practical, toward the rear edge of the work surface and away from the front grille of the cabinet.

¥ Place aerosol-generating equipment (e.g. vortex mixers, tabletop centrifuges) toward the rear of the cabinet.

¥ The workflow should be from “clean to dirty”. Materials and supplies should be placed in the cabinet in such a way as to limit the movement of “dirty” items over “clean” ones.

==Open Flames in a BSC:==
• Open flames in BSC are not to be used.

• If a researcher requests to use open flames, OSEH personnel will meet with the researcher and discuss issues and solutions.

• If it is deemed absolutely necessary for the work being done, use a pilotless burner or safety touch-plate microburners to provide a flame on demand. Refer to OSEH Advisory Use of Bunsen Burners in Biological Safety Cabinets for guidance.

• The University of Michigan has taken a strong stance against the use of gas burners or alcohol flames in BSC. This decision has been made in accordance with recommendations from numerous agencies. The Centers for Disease Control and Prevention (CDC) reports that “open-flames are not required in the near microbe-free environment of a biological safety cabinet” and create “turbulence which disrupts the pattern of air supplied to the work surface," jeopardizing the sterility of the work area. This is also the recommendation of the World Health Organization (WHO) as well as the major BSC manufacturers.

• Early microbiologists had to rely on open flames to ensure sterility. With the advancement of modern technology, including the introduction of the BSC, the use of an open flame is no longer necessary.

==UV Lights in a BSC:==
The Center for Disease Control (CDC) and the National Institutes of Health (NIH) agree that UV lamps are not recommended nor required in BSC. UV lamps must be turned off when the room is occupied to protect eyes and skin from UV exposure, which can burn the cornea and cause skin cancer. Proper use and cleaning of BSC negates any need for the use of UV lamps. Numerous factors affect the activity of the germicidal effect of UV light, which require regular cleaning, maintenance and monitoring to ensure germicidal activity.

=Protective Equipment=
Appropriate personal protective equipment (PPE) must be worn. Lab coats must be buttoned. Gloves should be pulled over the wrists of lab coat, not worn inside coat. Additional PPE to be used as recommended.

Note: Respirators are masks designed to protect the wearer from specific airborne hazards and are different from surgical masks, which protect the wearer only from splashes and are primarily intended to protect others from infectious aerosols exhaled by the wearer. Respirator use requires employee participation in the Respiratory Protection Program, which involves medical clearance and annual fit testing and training. Please be clear about use of surgical masks versus respirators. (Do NOT use the vague term “masks”).

=Waste Disposal=
A vacuum flask system is required to provide protection to the central building vacuum system or vacuum pump and to personnel who service the equipment. The figure below illustrates a proper set-up for handling liquid waste.

--missing picture--
The left suction flask (A) is used to collect the contaminated fluids into a suitable decontamination solution; the right flask (B) serves as a fluid overflow collection vessel. An in-line HEPA filter (C) is used to protect the vacuum system (D) from microorganisms.

• Connect the primary flask to an overflow collection flask and to an in-line HEPA filter.

• Both flasks shall contain an appropriate disinfectant for the material used.

• The vacuum flasks may be set up within the cabinet; however, to save room, the system can be placed on the floor beneath or next to the BSC, using a secondary container to contain the flasks and a longer hose connection to the vacuum system.

• Once inactivation occurs, liquid materials can be disposed of as noninfectious waste in the sink. Empty the waste from the flask when it reaches no higher than ¾ full. Replace the flask with fresh disinfectant.

Questions regarding waste pick up should be directed to OSEH Hazardous Materials Management (HMM) at (734) 763-4568. This office can also assist you in minimizing waste generation, providing waste containers, labels, and manifests.

=Exposures/Unintended Contact=

If the employee is in need of emergency medical attention, call 911 immediately.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm

After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021

University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320

UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

Click here for additional accident and injury reporting information.

=Spill Procedure=
• When a spill occurs, personal safety should always come first.

• Alert and clear everyone in the immediate area where the spill occurred.

A minor (small) biological spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., low concentration of infectious material that has high infectious dose small volume less than 1 Liter. A major/large biological spill, i.e., high concentration of infectious material that has low infectious dose, regardless of volume outside of BSC --may require active assistance from emergency personnel if volume is significant.

==Spill Response Steps:==

===MINOR or MAJOR BIOLOCIAL SPILL in BSC===
• BSC must be operating to provide user protection.

• Alert people in immediate area of spill.

• Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.

• Decontaminate all surfaces and items before removing from BSC.

• Cover spill with paper towel then saturate covered spill with disinfectant.

• Allow disinfectant 20 minutes of contact time before wiping up spill.

• Use tongs/forceps to pick up paper towel for disposal.

• Clean spill area with fresh towels soaked in disinfectant.

• Collect all waste materials in autoclave bag and autoclave using appropriate procedures.

• Clean spill area with water or 70% ethanol to preserve integrity of BSC.

• Allow cabinet to run an additional 10 minutes before resuming work or shutting down.

===MAJOR BIOLOGICAL SPILL outside BSC===
• Attend to injured or contaminated persons and remove them from exposure.

• Alert people in the laboratory to evacuate.

• Close doors to affected area.

• Post warnings to keep people from entering the area.

• Have person available that has knowledge of agents in use, incident, and laboratory to assist emergency personnel if assistance is requested.

• Wear PPE (gloves, lab coat, eye and respiratory protection).

• Cover spill with paper towel and saturate spill with disinfectant.

• Allow disinfectant 20 minutes of contact time before wiping up spill.

• Use tongs/forceps to pick up paper towel for disposal.

• Clean spill area with fresh towels soaked in disinfectant.

• Collect all waste materials in autoclave bag and autoclave using appropriate procedures.

Additional Spill Links:
• www.oseh.umich.edu/pdf/chemspil.pdf

• http://www.oseh.umich.edu/emer-chemical.shtml.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

=Training of Personnel=
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when working within a BSC.

=Certification=
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure.

*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 19:56, 10 November 2016 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:18, 12 June 2017 (UTC)

Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Principal Investigator? ☐ YES ☐ NO

Principal Investigator

SOP - Irritants

2017-06-12T22:15:02Z

Elhabbal: /* Certification */

[[Category: SOP]]
[[Category: Lab Safety]]

==Description==
This standard operating procedure outlines the handling and use of irritant chemicals. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling irritant chemicals.

A chemical irritant is one that is not corrosive but that causes a reversible inflammatory effect on living tissue by chemical action at the site of contact.

==Potential Hazards==
Irritants are chemicals that cause reversible inflammatory effects on living tissue by chemical action at the site of contact. A wide variety of organic and inorganic compounds are irritants; thus, skin contact with all laboratory chemicals should be avoided.
* Substances that may cause eye irritation include: YPD broth, Triton X-100, Trizma base, sodium pyrophosphate decahydrate, sodium fluoride, sodium dodecyl sulfate, poncaeu S, lithium chloride, Luria broth, glycerol, water-soluble dexamethasone, corn oil, and chloroform.
* Substances that may cause skin irritation include: Calcium chloride dehydrate, chloroform, corn oil, water-soluble dexamethasone, luria broth, lithium chloride, Meyer’s hematoxylin, limonene mount, ponceau S, sodium dodceyl sulfate, sodium pyrophosphate decahydrate, trizma base, and YPD broth.
* Substances that may cause respiratory irritation include: YPD broth, trizma base, sodium pyrophosphate decahydrate, sodium dodceyl sulfate, sodium deoxycholate, ponceau S, lithium chloride, luria broth, water soluble dexamethasone, corn oil, and calcium chloride dihydrate.

==Engineering Controls==
Use a properly functioning lab fume hood when handling irritants that can be inhaled (via mist/fume/gas/vapor). If the process does not permit the handing of such materials in a fume hood, contact Occupational Safety and Environmental Health (OSEH) at (734) 647-1143 to review the adequacy of ventilation measures.

==Work Practice Controls==
Handling processes should be designed to minimize the potential for splash, splatter, or other likely scenarios for accidental contact.

Ensure secondary containment and segregation of incompatible chemicals per guidance within the substance-specific storage guidance provided in SDS documentation.
To prevent contact with irritants, be sure to handle them carefully with the proper PPE.

For irritants that are also considered particularly hazardous substances, a designated area shall be established per other applicable SOPs.

==Personal Protective Equipment==
At minimum, safety glasses, lab coat, long pants, and closed toe shoes are to be worn when entering laboratories having hazardous chemicals.
Additionally:
* When handling hazardous chemicals or contacting potentially contaminated surfaces, protective gloves are to be worn. For proper selection of glove material, review the chemical-specific SDS.
* Goggles (not safety glasses) are appropriate for processes where splash or spray is foreseeable.
* For hazardous chemicals that are toxic via skin contact/ absorption, additional protective clothing (i.e., face shield, apron, oversleeves) is appropriate where chemical contact with body/skin is foreseeable.

==Transportation and Storage==
* Transport irritant chemicals in secondary containment, preferably a polyethylene or other non-reactive bottle carrier.
* Store in well-ventilated areas with secondary containment, such as a non-reactive plastic bin.
* Store below eye level.
* Store away from incompatibles. Review the chemical’s SDS for incompatibility information. Also refer to OSEH’s Chemical Storage webpage for more information.
* Avoid storing on the floor. If storing on the floor is necessary, use secondary containment.

==Waste Disposal==
Irritant substances intended for disposal may likely be considered hazardous wastes. Wherever possible, attempt to design research in a manner that reduces the quantity of waste generated. Contact OSEH’s Hazardous Materials Management (HMM) at (734) 763-4568 for waste containers, labels, manifests, and waste collection. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.

===For an actual chemical exposure/injury,===
* Flush exposed eyes or skin with water for at least 15 minutes, then seek medical attention (see below).
* Consult SDS for guidance on appropriate first aid. Where medical attention is required, be sure to bring along SDS(s) of chemical(s) to aid medical staff in proper diagnosis and treatment.
Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===
===U-M Occupational Health Services -- Campus Employees===
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
===University Health Services -- University students (non-life threatening conditions)===
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
===UMHS Emergency Department -- after clinic hours or on weekends===
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for more information.

==Spill Procedure==
Prompt response to chemical spills is critical to protect worker health & safety and to mitigate adverse effects to the environment. For further guidance, refer to chemical-specific MSDS.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., less than 1 gallon or 3.5 liters. A major/large chemical spill requires active assistance from emergency personnel.

===MINOR CHEMICAL SPILL===
* Alert people in immediate area of spill.
* If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
* Open outside windows, if possible.
* Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.
* Avoid breathing vapors from spill.
* Confine spill to as small an area as possible.
* Do not wash spill down the drain.
* Use appropriate spill kits/sorbents to absorb spill. Collect contaminated materials and residues and place in container. For powdered chemicals sweep carefully to avoid generation of dust or, if appropriate, use moist sorbent pads or wet the powder with a suitable solvent and then wipe with a dry cloth. Contact OSEH-HMM (734) 763-4568 for proper disposal.
* Clean spill area with water.

===MAJOR CHEMICAL SPILL===
* Attend to injured or contaminated persons and remove them from exposure.
* Alert people in the laboratory to evacuate.
* If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
* Call University of Michigan Division of Public Safety and Security (DPSS) at 911 immediately for assistance.
* Close doors to affected area.
* Post warnings to keep people from entering the area.
* Have person available that has knowledge of incident and laboratory to assist emergency personnel.

Additional Spill Links:
* www.oseh.umich.edu/pdf/chemspil.pdf
* http://www.oseh.umich.edu/emer-chemical.shtml.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling irritant chemicals.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:27, 10 November 2016 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 16:14, 31 October 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:52, 7 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:05, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:15, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO
Laboratory Director - Dave Bridges Revision Date - 2016-10-19

SOP - Cryogenic Materials

2017-06-12T22:11:25Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety]]
=Description=
This standard operating procedure (SOP) outlines the handling and use of cryogenic materials. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling cryogenic materials.

Cryogenic liquids have various uses in the laboratory including sample preservation, cooling of equipment, and handling and storage of biological samples.

=Potential Hazards=
Cryogenic materials are liquefied gases with boiling points less than -73°C (-100°F). They can cause tissue damage (frostbite) and asphyxiation due to oxygen displacement. They also pose a fire hazard if oxygen is condensed out of the atmosphere during the use of cryogenic liquids that have boiling points that are lower than oxygen – e.g. nitrogen, helium or hydrogen. There is a potential for explosion due to pressure buildup in closed containers. Materials intended for use with cryogenic liquids must be selected carefully as they can be weakened or fail due to embrittlement.

=Engineering Controls=
Use and store cryogens in well-ventilated areas. Closets, small rooms, and walk-in cold rooms (or similar small/unvented spaces) should be avoided to prevent buildup of the gas as the cryogen evaporates/sublimes. If using large quantities of cryogens or if there are concerns about the adequacy of ventilation, consult OSEH Research Health and Safety (734) 647-1143 to determine if additional ventilation or an oxygen sensor/alarm is needed.

Each part of a cryogenic system must have its own pressure relief system.

=Work Practice Controls=
• Cryogenic gases can condense oxygen out of the air, creating a localized oxygen enriched environment. Therefore they must be used and stored away from flammable materials, open flames and ignition sources.

• Do not put your head inside a liquid nitrogen freezer or other enclosed space containing a cryogen. Never allow people to travel in an elevator with cryogenic liquids.

• Do not use hollow rods or tubes as dipsticks. (When a warm tube is inserted into a cryogen, liquid will spout from the top of the tube.

• When retrieving cryopreservation vials that have been stored in liquid nitrogen, raise the vials out of the liquid phase and store them in the gas above the liquid for 24 hours to allow any trapped liquid nitrogen to escape. After this period, the vials may be removed. Retrieving these vials directly from the liquid phase may cause the vials to rupture if liquid nitrogen has become trapped inside. This may expose the employee to flying bits of plastic, biological specimens or cryoprotective agents (such as dimethyl sulfoxide) that were in the vial.

• Check the liquid levels of Dewars regularly. If the liquid evaporates more rapidly than normal, the Dewar may be losing its vacuum.

• Also follow the SOP for asphyxiants.

=Personal Protective Equipment (PPE)=
• Wear loose fitting cryogenic gloves, safety goggles, face shield, lab coat, long pants and closed toe shoes when handling and transporting cryogenic materials.

• Lab coats should be knee length and have no pockets or cuffs to catch the liquid.

• Pants should not have cuffs and should be long enough to go over the shoe or boot.

=Transportation and Storage=
• Never store cryogenic liquids or dry ice in walk-in cold rooms.

• Use and store liquid hydrogen and helium away from flammable materials and ignition sources. (These gases can condense oxygen out of the air, creating a localized oxygen enriched environment.)

• For liquid helium and hydrogen storage systems, check the pressure relief and inspect the system for leaks regularly.

• These gases are cold enough to solidify atmospheric air; leaks in storage systems for these gases may become plugged with solidified air. If the pressure relief device becomes plugged, the container may over-pressurize and fail.

• Use and store liquid oxygen away from open flames. Post a “No Open Flames” sign in liquid oxygen storage areas.

• Store and transport cryogenic materials only in Dewars or cryogenic liquid cylinders designed for that particular cryogen. Inspect Dewars daily to insure that no air or ice plugs exist in the neck openings.

• Cryogens may be transported in elevators only in containers certified to leak at less than or equal to 1 liter of liquid (or 1 kilogram of solid) per day. Never allow passengers to travel in an elevator with cryogenic liquids.

=Waste Disposal=
If a cryogen (such as dry ice) has been mixed with a liquid that would normally be disposed of as hazardous waste (such as alcohol, as in a cold trap), follow disposal procedures for that liquid. • Cryogens must not be dumped into sinks, as the thermal shock is likely to damage the sink and/or the plumbing.

• As the unneeded cryogen is evaporating, good ventilation is essential to prevent hazardous air concentrations of the gas.

• Do not put unneeded cryogen in a sealed container, as the build-up of pressure could cause an explosion.

=Exposures/Unintended Contact=

If the employee is in need of emergency medical attention, call 911 immediately.

For an actual chemical exposure/injury:
• Refer to SDS.

• Flush exposed eyes or skin with water with warm water for at least 15 minutes, then seek medical attention (see below).

• Do not rub affected area.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to Work Connections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the Work Connections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm

After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021

University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320

UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

=Spill Procedure=
• When a spill occurs, personal safety should always come first.

• Alert and clear everyone in the immediate area where the spill occurred.

Spills of cryogenic liquid can cause depletion of oxygen in the area. For spills over 1 liter (or smaller amounts in very small rooms), leave the area immediately and call the University of Michigan Division of Public Safety and Security (DPSS) at 911.
Additional Spill Links:
• http://oseh.umich.edu/hazardous-waste/spill-response/

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

=Training of Personnel=
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling cryogenic materials.

=Certification=
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure.

*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 20:44, 10 November 2016 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:11, 12 June 2017 (UTC)

Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☐ NO

Laboratory Director Revision Date
�
Major Revisions (Tracking purposes only – Do not print as part of SOP)

SOP - Autoclave

2017-06-12T22:09:41Z

Elhabbal: /* Certification */

[[Category: SOP]]
[[Category: Lab Safety]]

==Description==
This standard operating procedure outlines the use of autoclaves
Autoclaves are used to sterilize equipment through a process that incorporates high pressure and high temperature.

==Potential Hazards==
Autoclaves pose several potential hazards to labs who frequently use them. Among the physical hazards are heat, steam, and high pressure. Biological hazards, including improperly autoclaved infectious materials, are also present.

Controls for different brands of autoclaves may have their own unique characteristics for loading, load sizes, cycle types and cycle settings. The type of materials to be sterilized will determine the sterilization cycle that will be used. For this reason it is important to read and understand the user’s manual for the specific model of autoclave that is being used prior to first time use. Always ensure that the user’s manual is readily available in case questions or concerns arise during the operation of the autoclave.

==Engineering Controls==
Autoclave monitoring and maintenance is an important aspect of a safe and properly functioning autoclave. Follow the manufacturer’s recommendations for preventive maintenance and ensure that all contractors hired to perform regular maintenance and repairs are approved by the manufacturer. Autoclave operators shall ensure that each autoclave is monitored as follows:

Heat Sensitive Tape Monitoring – Operators shall use heat sensitive sterilization indicator tape for each load to indicate that the load has undergone an effective steam sterilization process.
* Change on autoclave tape means the proper temperature was achieved, but does not mean that temperature was maintained for the proper length of time.
* Place a small piece of autoclave tape on each bottle and tip box.
Biological Indicators – Operators who autoclave medical/biohazardous waste need to do the following:
* At least once a month use a biological indicator such as Bacillus Stearothermophilus (e.g. Prospore2) placed at the center of a load processed under standard operating conditions to confirm the attainment of adequate sterilization conditions.
==Work Practice Controls==
Do not autoclave items containing corrosives, solvents, volatiles or radioactive materials.

===Prior to loading===
* Before using the autoclave, be sure previous users have cleared the autoclave, and that no broken items have been left inside.
* Check to be sure the drain is clear.
* Verify that the interior of the autoclave is in usable condition.

===Loading===
* Carefully load items to be autoclaved, DO NOT overload the autoclave.
* Bottles containing liquids should be placed inside a heat resistant plastic tray (secondary container) containing an inch of water.
* Bottles should not be filled more than 2/3.
* Keep 1-2 inches of space between bottles.
* DO NOT tighten caps on bottles fully.
* Individual glassware pieces should be within a secondary container on a shelf or rack and never placed directly on the autoclave chamber bottom or floor.
* Make sure that the door of the autoclave is fully closed and latched and ensure that the correct cycle for the items being autoclaved has been selected before starting the cycle.

===Opening===
* Wear the proper PPE, including autoclave gloves and close toed shoes when opening the autoclave door after a cycle.
* When the cycle is complete, open the door slowly to minimize exposure to steam. Keep appendages away from the opening.

===When autoclaving liquids:===
* When running an autoclave cycle with liquids, choose the liquid cycle setting.
* To prevent bottles from shattering during the pressurization, the caps of containers with liquids must be loosened before loading.
* Use only borosilicate glass (Pyrex™ or Kimax™).
* Use a secondary container with a solid bottom and walls to contain the contents and catch spills.
* Set a timer to come and unload the autoclave, to ensure it is available for use by other labs.
* Wait 10 minutes after the cycle ends before removing autoclaved liquid load items.
* Let the liquids stand for at least a full hour before touching with ungloved hands. Be sure to let others in the area know that a heat hazard is present.

===When autoclaving dry loads:===
* Add ¼ to ½ inch of water to the tray so that the bottles will heat evenly. Do not fully tighten bottle caps, this could lead to the bottle shattering. Loose caps allow in inside of the bottle to be sterilized.
* Check plastic materials to ensure that they are compatible with being autoclaved.
* Choose the dry porous materials cycle.
* Set a timer to come and unload the autoclave, to ensure it is available for use by other labs.
* Before removing autoclaved items, wait 5 minutes after the cycle ends for loads containing only dry glassware.
* For dry loads, let the glassware cool for a minimum of 15 minutes before touching it with ungloved hands.

==Protective Equipment (PPE)==
Wear the proper PPE, including heat resistant gloves, and close toed shoes when opening the autoclave door after a cycle.
==Waste Disposal==
Place waste in a university approved autoclave bag; avoid over packing of autoclave bags. Do not seal bags or close bottles and other containers tightly. After sterilization cycle, close and seal bag. The sterilized waste can be transported directly to the dumpster by Building Services or laboratory staff without boxing or labeling. Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.

Discontinue use immediately if an autoclave is not working properly. Post a sign alerting others not to use the autoclave. Mechanical failures need to be attended to by a trained technician, DO NOT try to fix it yourself. Contact facilities to have it repaired.
If you are burned, you should seek medical treatment immediately. Burns to the face, third- degree burns or burns over large areas of the body should be treated as emergencies. (Call 911). Minor burns should be treated by using first aid procedures, including immersing the burn in cool water immediately, removing clothing from the burn area and keeping the injured area cool for at least 5 minutes. Regardless of the severity, notify your Laboratory Director/Supervisor!

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

====TREATMENT FACILITIES:====
====U-M Occupational Health Services -- Campus Employees====
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
====University Health Services -- University students (non-life threatening conditions)====
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
====UMHS Emergency Department -- after clinic hours or on weekends====
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for more information.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of personnel==
All personnel are required to complete the Autoclave Safety training online through MyLINC (Course BLS013w). All users shall be trained before operating an autoclave; the Laboratory Director/Supervisor is responsible for ensuring that each person in the lab is appropriately trained. All training must be documented and the records maintained in the lab with other safety training certificates. The Laboratory Director/Supervisor is encouraged to use this policy as a guide for training new personnel.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 16:13, 19 October 2016 (UTC)
* [[User:Davebridges|Davebridges]] ([[User talk:Davebridges|talk]]) 14:53, 8 November 2016 (UTC)
*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:11, 5 June 2017 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:43, 7 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:04, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:09, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? X YES ☐ NO
Laboratory Director - Dave Bridges Revision Date - 2016-10-19

SOP - Chloroform

2017-06-12T22:06:55Z

Elhabbal: /* Certification */

[[ Category: SOP ]]

==Description==
This standard operating procedure outlines the handling and use of chloroform. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling chloroform.

Chloroform (CAS # = 67-66-3) is a clear, colorless liquid with a pleasant, nonirritating odor that can be detected as low as 11.7 ppm (odor threshold) but more likely detected at levels of 85 to 307 ppm (well above safe exposure limits). It will burn only when it reaches very high temperatures.

NOTE: Stabilizers are often added to prevent oxidation by air and light, and to prevent chloroform from becoming acidic and corrosive.

Synonyms include: Trichloromethane, Methane trichloride, Formyl trichloride, Trichloroform, Methenyl trichloride and Freon 20.

Chloroform is used as an extractant solvent in manufacture of rubber, essential oils, sterols and alkaloids, guttapercha, resins, and in the recovery of fat from waste products; in chemical analysis and assays; and in photographic processing.
It is also used as a general solvent of lacquers, plastics, dyes, fats, greases, gums, oils, adhesives, and waxes, and in the rubber cleaning and dry cleaning industries.

==Useful Chloroform Links:==
* http://www.atsdr.cdc.gov/substances/toxsubstance.asp?toxid=16
* http://www.atsdr.cdc.gov/toxfaqs/tf.asp?id=52&tid=16
* http://www.nap.edu/openbook.php?record_id=4911&page=282

==Potential Hazards==
* Chloroform has been classified as a potential to probable human carcinogen, based on adequate evidence for carcinogenicity in animals. Some animal studies also show evidence of reproductive and developmental toxicity from chloroform exposure.
* Inhalation of vapors can cause headaches, drowsiness, dizziness, and nausea. At high concentrations disorientation, anesthetic effects, and unconsciousness can occur, but acute toxicity is low.
* Chloroform is an eye, skin and upper respiratory tract irritant.
* Chloroform is not combustible but exposure to fire or high temperatures may lead to formation of phosgene, hydrogen chloride and chlorine, all highly toxic gases.
* Consult the SDS for Chloroform for additional guidance and information.

Occupational Exposure Limits (OELs):
* MIOSHA: 2 ppm, 8-hour PEL

==Engineering Controls==
Work with chloroform in a chemical fume hood. Dilute solutions (< 10%) may be used on the benchtop in small quantities (< 500 mL).

==Work Practice Controls==
* Keep containers of chloroform closed as much as possible.
* Be aware of skin absorption as a possible route of exposure. Plan work so that minimal glove contact is expected, and purchase appropriate gloves for cleaning up small spills. (Refer to the PPE section below, for glove recommendations.)
* Use in the smallest practical quantities for the experiment being performed.
* Do not mix or store with acids; may form toxic gas.
* Thoroughly wash hands when finished handling.

==Personal Protective Equipment (PPE)==
* The minimum PPE for work with chloroform is Viton or PVA (Polyvinyl Acetate) laboratory gloves, lab coat, and safety glasses (ANSI/ISEA Z87.1 approved). Keep in mind that chloroform will penetrate nitrile gloves in less than 2.5 minutes.
* If a splash may occur, wear chemical splash goggles and/or a face shield.
* OSEH’s Glove Compatibility webpage can also be used to determine the recommended gloves.

==Transportation and Storage==
* Do not store chloroform with incompatibles. Chloroform is not compatible with the following: acetone, strong bases, alkalis, chemically-active metals (such as aluminum, magnesium, sodium, or potassium), dinitrogen tetroxide, fluorine, disilane, sodium methylate, triisopropylphosphine, and solid potassium tert-butoxide.
* Transport chloroform in secondary containment, preferably a polyethylene or other non-reactive acid/solvent bottle carrier.
* Store in secondary containment.
* Avoid storing on the floor.
* Refer to OSEH’s Chemical Storage webpage for additional information on the storage of hazardous materials.

==Waste Disposal==
Because most spent, unused and expired chemicals/materials are considered hazardous wastes, they must be properly disposed of. Do not dispose of chemical wastes by dumping them down a sink, flushing in a toilet or discarding in regular trash containers, unless authorized by OSEH Hazardous Materials Management (HMM). Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.
* Eyes: In case of contact, immediately flush eyes with plenty of water for at least 15 minutes. Get medical aid.
* Skin: In case of contact, immediately flush skin with plenty of water. Remove contaminated clothing and shoes. Get medical aid. Wash clothing before reuse.
* Ingestion: Get medical aid immediately. Do not induce vomiting unless directed to do so by medical personnel. Never give anything by mouth to an unconscious person. If vomiting occurs naturally, have victim lean forward.
* Inhalation: If inhaled, remove to fresh air. If not breathing, give artificial respiration. If breathing is difficult, give oxygen. Get medical aid.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for more information.

==Spill Procedure==
* When a spill occurs, personal safety should always come first.
* Alert and clear everyone in the immediate area where the spill occurred.
* Use proper personal protective equipment (PPE) as noted above.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., (less than 1 Gallon or 3.5 Liters). A major/large chemical spill requires active assistance from emergency personnel.

===Spill Response Steps:===

===MINOR CHEMICAL SPILL===
* Alert people in immediate area of spill.
* Open outside windows, if possible.
* Use proper personal protective equipment as indicated above.
* Avoid breathing vapors from spill.
* Confine spill to as small an area as possible.
* Do not wash spill down the drain.
* Use appropriate spill kits/sorbents to absorb spill. Collect contaminated materials and residues and place in container. Contact OSEH-HMM (734) 763-4568 for proper disposal.
* Clean spill area with water.

===MAJOR CHEMICAL SPILL===
Report large chemical spills greater than 1 Gallon or 3.5 Liters in corridors or common areas, e.g., hallways, elevators, eating areas, rest rooms, offices, etc., to U-M Division of Public Safety and Security (DPSS) by calling 911.
* Attend to injured or contaminated persons and remove them from exposure.
* Alert people in the laboratory to evacuate.
* Call U-M Division of Public Safety and Security (DPSS) at 911 immediately for assistance.
* Close doors to affected area.
* Post warnings to keep people from entering the area.
* Have person available that has knowledge of incident and laboratory to assist emergency personnel.

Additional Spill Links:
* www.oseh.umich.edu/pdf/chemspil.pdf
* http://www.oseh.umich.edu/emer-chemical.shtml

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling chloroform.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Davebridges|Davebridges]] ([[User talk:Davebridges|talk]]) 14:42, 15 August 2016 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 16:15, 31 October 2016 (UTC)
*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:09, 5 June 2017 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 18:50, 7 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:01, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:06, 12 June 2017 (UTC)

Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO

Laboratory Director - Dave Bridges Revision Date - 2016-08-15

SOP - Compressed Gases

2017-06-12T22:04:59Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
This standard operating procedure outlines the handling and use of compressed gases. Compressed gases come in a large variety of sizes and pressures. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling compressed gases.

A compressed gas is any mixture or material in a container with either an absolute pressure exceeding 40 psi at 70F or an absolute pressure exceeding 104 psi at 130F. Any liquid flammable material having a vapor pressure exceeding 40 psi at 100F is also considered a compressed gas.

==Potential Hazards==
The large amount of potential energy contained in a compressed gas cylinder makes it a potential rocket or bomb if the pressure is released through rupture of the valve or container failure.

Compressed gases may also be toxic, flammable, or explosive – check the safety data sheet for more information. Safety considerations for these properties must also be followed.

==Engineering Controls==
Storage of compressed gas cylinders requires sturdy anchoring to a wall or cabinet, and/or a cylinder stand. If the process does not permit gas use and/or storage in well-ventilated areas (i.e., lab ventilation having a minimum of 6 air changes per hour), contact Occupational Safety and Environmental Health (OSEH) at (734) 647-1143 to determine the necessity of an oxygen-deficiency monitor or other alarm devices.

W==ork Practice Controls==
All compressed gas cylinders shall be legibly marked by stenciling, stamping, or label with at least the chemical name or commonly accepted name of the material contained. In addition, cylinders should bear the approved markings of the Department of Transportation stamped in the metal at the top of the cylinder.
* Check connections and hoses near the cell culture room and the animal procedure room regularly for leaks using a specific monitoring instrument or soapy water (or equivalent). Some leaks can be heard/felt during use.
* Be sure that cylinders are properly labeled as full, in use, or empty.
* When using highly flammable or toxic gas, check the delivery system using an inert gas prior to introducing the hazardous gas.
* When using compressed acetylene: (i) do not exceed a working pressure of 15 psig, and (ii) do not use vessels, piping, or other materials that contain a significant amount of copper (usually considered to be more than 50% copper).
* Replace valve caps when cylinders are not in use or before moving.
* Remove damaged or defective cylinders from service (contact the cylinder vendor for assistance).
* Remove unused or empty cylinders from lab space. Replace them with full cylinders.
* Be sure to fully close cage lid before turning on CO2.
* Refer to the OSEH Compressed Gas Guideline or consult with your OSEH representative regarding maximum allowable quantities of compressed gases.

Restricted hazardous gas use must be approved by OSEH for purchase. Refer to the hazard guideline for compressed gas use on the OSEH website or contact your OSEH representative for more information. This form can be used to submit a request for approval of the purchase of a restricted gas.

==Protective Equipment==
Safety glasses must be worn for all work involving compressed gas cylinders. Cylinders must be secured to a gas cylinder mount, bracket, or clamp. These securing devices must be attached to a stable surface such as a permanent bench top or a wall.

==Transportation and Storage==
* Cylinders (full, in use, or empty) shall be secured by chains, straps, or other sturdy tie downs during storage and transport.
* Cylinders shall be grouped by type of gas, and the groups segregated as to compatibility.
* Full cylinders shall be separated from empty cylinders within the storage area.
* Flammable gases shall be separated from nonflammable gases by at least 20 feet.
* Cylinders shall not be stored at temperatures above 125 F. or in direct sunlight, or outside of the temperature range specified by the manufacturer.
* Cylinder valves shall be kept closed when not in use.
* Removable caps shall be kept on cylinders at all times, unless they are in use.
* Cylinders shall be protected against tampering and damage.
* Cylinders shall not be stored near combustible materials.
* Cylinders shall not be refilled except by authorized suppliers.
* Open flames and smoking shall not be permitted in areas where oxygen is used or stored. "No Smoking" and "No Open Flames" signs shall be conspicuously posted in these areas.
* Cylinders, except for those containing compressed air, shall not be used or stored in cold rooms or other unventilated enclosures. An exception may be approved by OSEH for inert gases when an oxygen monitor is in place.

==Waste Disposal==
In most cases, the compressed gas cylinder, including any unused gas, will be returned to the vendor from which the cylinder was purchased. Contact Occupational Safety and Environmental health (OSEH) Hazardous Materials Management (HMM) at 734-763-4568 to arrange for the removal of gas cylinders that cannot be returned to the supplier. Write “empty” on the outside of each cylinder and complete a hazardous waste manifest.
==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
===University Health Services -- University students (non-life threatening conditions)===
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
===UMHS Emergency Department -- after clinic hours or on weekends===
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for more information.

==Release/Spill Procedure==
Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling and using compressed gases.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO

*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 15:55, 31 October 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:25, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:44, 7 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:04, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:04, 12 June 2017 (UTC)

Laboratory Director - Dave Bridges Revision Date - 2016-10-19

SOP - Cytotoxins and Antineoplastic Drugs

2017-06-12T22:02:45Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety]]

'''Principal Investigator (PI) Approval is Required Prior to Performing this Procedure'''
=Description=
This standard operating procedure outlines the handling and use of cytotoxic chemicals and antineoplastic drugs. Review this document and supply the information required in order to make it specific to your facility. In accordance with this document, laboratories should use appropriate controls, protective equipment, and disposal techniques when handling cytotoxic chemicals and antineoplastic drugs.

=Potential Hazards=
Although little research has been done on the long-term risk of occupational exposure to cytotoxic drugs, these drugs have been associated with human cancers at high (therapeutic) levels of exposure. These drugs have been shown to be carcinogens, mutagens and teratogens in many animal species. There is evidence that hazardous drugs may cause spontaneous abortions and increase the risk of congenital malformations. In addition, some of these drugs have been shown to cause acute effects in humans, such as localized skin necrosis (death of tissue), damage to normal skin after surface contact, dizziness, lightheadedness.

Antineoplastic drugs are substances used as a cancer chemotherapeutic agent which affect the reproductive capabilities including chromosomal damage (mutagens), effects on the fetus (teratogens), and are carcinogenic in animal models. Most hazardous drugs either bind directly to genetic material in the cell nucleus or affect cellular protein synthesis. Cytotoxic drugs may not distinguish between normal and cancerous cells. The growth and reproduction of the normal cells are often affected during treatment of cancerous cells.

=Engineering Controls=
All hazardous drug work will be conducted in a chemical fume hood or Class II Biological Safety Cabinet.

=Work Practice Controls=
Decontamination should consist of surface cleaning with water and detergent followed by thorough rinsing. The use of detergent is recommended because there is no single accepted method of chemical deactivation for all agents involved. A plastic backed absorbent pad should be placed under the work area during the process. This should be changed at the end of each process or when a spill occurs.

All locations within the laboratory where cytotoxic or antineoplastic chemicals are handled should be demarcated with designated area caution tape and/or posted with designated area caution signs. This includes all fume hoods and bench tops where the antineoplastic drugs are handled.

=Personal Protective Equipment=
Use doubled gloves with nitrile or neoprene gloves. Gloves should be changed frequently. Laboratory coat and safety glasses are required for preparation of material.
Laboratory SOP: Cytotoxins 1 Revision Date: 6/12/2014

=Waste Disposal=
All hazardous drug contaminated waste should be placed in a 5-gallon white pail. The container must be closed except when actively adding waste. The container must be located in the area where hazardous drugs are being used. All items contaminated by hazardous drugs, including gloves, syringes, vials, needles, and solution containers should be disposed according to Environment, Health and Safety (EHS) guidelines. Contact EHS at (810) 766-6763 for waste pick-up. Refer to UM-Flint Hazardous Waste Management Program and EHS webpage http://www.umflint.edu/ehs/environment- health-and-safety for more information.
Sharps – place needles, syringes with needles attached and other breakable items into appropriately labeled sharps containers.
Empty stock vials, reagent bottles, etc. – triple rinse with copious amounts of water. Deface label with black magic marker or scraper. Place in a cardboard box for disposal. Label the box as uncontaminated glass.

=Exposures/Unintended Contact=

For an actual chemical exposure/injury:
• Flush exposed eyes or skin with water for at least 15 minutes, then seek medical attention (see below).

• Note: In case of inhalation, symptoms may be delayed up to 24 hours. Seek medical attention.

• If swallowed, obtain urgent medical attention.

Contact EHS for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.
add here...
===TREATMENT FACILITIES:===

=== MAJOR INJURIES===
'''Genesys Hospital'''
One Genesys Parkway Grand Blanc, MI 48439 (810) 606-5710

'''Hurley Medical Center'''
One Hurley Plaza Flint, MI 48503 (810) 262-9000

'''McLaren Hospital Flint'''
401 South Ballenger Hwy Flint, MI 48532
(810) 342-2000
===MINOR INJURIES –During Business Hours===
'''Genesys Occupational Health Network'''
1460 Center Rd.
Burton, MI 48509
(810) 715-4620
Mon. to Fri. 7:30 am to 10 pm Sat. & Sun. Noon to 8 pm

'''McLaren Flint-Burton OCC Center'''
1459 S. Center Rd. Burton, MI 48509
(810) 496-0900
Mon. - Fri. 8 am to 8 pm Sat & Sun 10 am to 2 pm

===MINOR INJURIES –After Business Hours===
'''Downtown Flint'''
420 S. Saginaw St. Flint, MI 48502 (810) 762-1550

'''Genesys East'''
1096 S. Belsay Rd, Suite F Burton, MI 48509
(810) 743-3351

'''Genesys North'''
4154 W. Vienna Rd Clio, MI 48420 (810) 686-7397

'''Genesys South'''
8447 N. Holly Rd
Grand Blanc, MI 48439
(810) 603-0856
Mon. - Fri. 6 to 10pm / Sat. & Sun. 1-10pm

Click here for more information on the UM – Flint Emergency Preparedness and Response Plan.

=Spill Procedure=
• When a spill occurs, personal safety should always come first.

• Alert and clear everyone in the immediate area where the spill occurred.

A minor (small) chemical spill is one of a known chemical that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., less than 1 Gallon or 3.5 Liters. A major/large chemical spill requires active assistance from emergency personnel.
'''Spill Response Steps:'''
===MINOR CHEMICAL SPILL===
• Alert people in immediate area of spill.

• If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other
switches.

• Open outside windows, if possible.

• Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.

• Avoid breathing vapors from spill.

• Confine spill to as small an area as possible.

• Do not wash spill down the drain.

•Use appropriate spill kits/sorbents to neutralize corrosives and/or absorb spill. Collect contaminated materials and residues and place in container. For powdered chemicals sweep carefully to avoid generation of dust or, if appropriate, use moist sorbent pads or wet the powder with a suitable solvent and then wipe with a dry cloth.

Contact EHS at (810) 766-6763 for proper disposal.

• Clean spill area with water.

===MAJOR CHEMICAL SPILL===
• Attend to injured or contaminated persons and remove them from exposure.

• Alert people in the laboratory to evacuate.

• If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.

• Call UM-Flint DPS at 911 from any university telephone or (810) 762-3333 from any cell phone or non- university telephone immediately for assistance.

• Close doors to affected area.

• Post warnings to keep people from entering the area.

• Have person available that has knowledge of incident and laboratory to assist emergency personnel.

Additional Spill Links:

• www.oseh.umich.edu/pdf/chemspil.pdf

• http://www.oseh.umich.edu/emer-chemical.shtml.

Report all emergencies, suspicious activity, injuries, spills, and fires to the UM-Flint Department of Public Safety (DPS) at 911 from any university telephone or (810) 762-3333 from cell phone or non-university telephone. Register with the University of Michigan-Flint Emergency Alert System via Wolverine Access. Also, preprogram the UM-Flint DPS telephone number (810) 762-3333 into your cell phone for quick, easy use.

=Training of Personnel=
All personnel are required to complete Laboratory Safety Training. Documentation of the training is required. This training can be accomplished by completing the Comprehensive Laboratory Safety session (BLS009 or equivalent) via MyLINC, or UM-Flint EHS on-line training or other equivalent approved by EHS. Furthermore, all personnel shall read and fully adhere to this SOP when handling cytotoxins and antineoplastic drugs.

=Certification=
I have read and understand the above SOP. I agree to contact my Supervisor or Lab Manager if I plan to modify this procedure.

*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:12, 10 November 2016 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:02, 12 June 2017 (UTC)
Principal Investigator Revision Date
Laboratory SOP: Cytotoxins 4 Revision Date: 6/12/2014

SOP - Needle Capping

2017-06-12T22:00:53Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
This procedure outlines the methods for the safe recapping of needles and covers the safe handling, practices, disposal, and needlestick reporting requirements pertaining to hypodermic needles, hypodermic needles with syringes, needles with attached tubing, and other needle equipment. It is the responsibility of all laboratory staff to contribute to and comply with these procedures.

==Potential Hazards==
Needles present serious health hazards because people can be easily exposed parenterally to hazardous chemicals and infectious biological agents through injection. Recapping needles is extremely dangerous because it can result in the accidental stabbing of hands and other body parts. If working with human blood or other potentially infectious material, ensure that an Exposure Control Plan is completed and available. All applicable work requirements such as SOPs, universal precautions, hepatitis vaccination, etc. are required to be followed.

==Engineering Controls/Workplace Controls==
NEEDLES SHOULD NOT BE RECAPPED, BENT, REMOVED OR OTHERWISE MANIPULATED BY HAND.

* However, if it is essential that a needle be recapped due to the nature of the work, the use of a mechanical device or the one-handed scoop method must be used. In addition, there needs to be written justification and instructions for any procedures that involves the recapping of needles (SOP).

==Needle Recapping Instructions:==
1. One-Handed Scoop Method

Step 1: Place the cap on the desk or other flat surface with something firm to “push” the needle cap against.

Step 2: Holding the syringe with needle attached in one hand, slip the needle into the cap without using the other hand.

Step 3: Push the capped needle against a firm object to “seat” the cap onto the needle firmly using only one hand.

2. Needle Recapping Devices

These are devices with a hole in the center that hold needle caps and allow for single hand uncapping and recapping.

==Additional Guidelines==
* Keep needles away from fingers and other body parts.
* Place uncapped needles in a rigid tray during procedures and transportation.
* Do not leave unprotected needles on work surfaces.
* Shearing or breaking of contaminated needles is prohibited.
* Use disposable needle locking syringe units and other engineered sharps injury protections, which has a built-in safety feature or mechanism that effectively reduce exposures, and needleless systems whenever possible.
* Blunt cannulas should be used as alternatives to needles wherever possible.
* Needles and syringes should never be used as a substitute for pipettes.
* Used disposable needles and syringes must be placed in conveniently located appropriate sharps disposal containers and discarded as infectious waste.
* Use extreme caution and be alert when handling sharps. Avoid distractions and keep sight of the needle.
* Ensure that animals are restrained when using needles on animals and be on the alert for any unexpected movements.
* The use of needles and syringes should be restricted to procedures for which there is no alternative.
* Refer to the OSEH Exposure Control Plan and Biosafety Manual for additional information.

==Personal Protective Equipment (PPE)==
Wear safety glasses/goggles, gloves, lab coat/gown, close-toe shoes, and any other applicable PPE when handling needles and syringes.

==Waste Disposal==
Upon completion of a procedure involving use of needle equipment, immediately discard the items, including syringes connected to the needle, tubing, etc., into a clearly and properly labeled or color-coded rigid, closable, leakproof, puncture-resistant, disposable sharps container located in the work area at the point of use. All used needle equipment waste must be discarded as infectious waste and in compliance with OSEH requirements. NEVER reach into the sharps container. Never empty the contents into another container. Do not overfill the sharps container.

==Accidental Needle Stick/Exposures/Unintended Contact==
If the employee is in need of emergency medical attention, call 911 immediately.

If a needlestick injury should occur, wash the puncture area with soap and water, report the incident to the supervisor and principal investigator, and seek medical attention as soon as possible. Any needlestick with recombinant DNA and/or an infectious agent must be reported to the U-M Biological Safety Officer (BSO) (763-3133) as soon as possible. The U-M BSO will determine if institutional notification to Federal Agencies is required within 24 hours per applicable regulations. Failure to report may result in severe penalties to the researcher, laboratory, department or institution.
Report all needlesticks to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Please include (1) the type and brand of device involved in the incident. (2) The work unit or work area where the exposure incident occurred. (3) An explanation of how the incident occurred. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

In addition, as required for all lab incidents, complete the OSEH Laboratory Incident and Near-Miss Report form.

TREATMENT FACILITIES:
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for additional accident and injury reporting information.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling needles.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab Manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Ccousz|Ccousz]] ([[User talk:Ccousz|talk]])
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:24, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:33, 7 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:03, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:00, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☒ NO

Laboratory Director - Dave Bridges Revision Date - 2016-10-14

SOP - Centrifuge Safety

2017-06-12T21:59:09Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
This standard operating procedure outlines the use of centrifuges. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls and personal protective equipment when using centrifuges.

Centrifuges are machines used to separate solids from liquids in a suspension. The spinning motion of a centrifuge produces centrifugal forces that separate substances of greater and lesser densities.

==Potential Hazards==
The centrifuge must be loaded carefully balanced. An unbalanced load may present a risk to both the machine and to persons nearby. Centrifuge rotors should never be touched while the rotor is spinning, as they present a significant personal risk when in motion. Another potential hazard is the possible aerosolization of harmful samples during centrifuge operation.

==Engineering Controls==
If centrifuging hazardous materials, care should be taken to use tightly capped tubes and/or sealable safety cups or rotors that can be loaded and unloaded in a fume hood or biosafety cabinet, depending on the hazard.

==Work Practice Controls==
The following information may be integrated into a lab-specific standard operating procedure (SOP) for centrifuge use.

1. ''Preventive Maintenance''

A. Establish preventive maintenance schedule: Including regular cleaning of centrifuge interior to prevent damage and avoid costly repairs. Reference centrifuge operator’s manual or contact manufacturer for guidance.

Equipment repair and adjustments shall only be conducted by qualified service technicians.

B. Maintain log book: For all high speed centrifuges and ultracentrifuges include run dates, durations, speeds, total rotor revolutions, and notes on rotor condition.

Retire rotors after manufacturer’s recommended life span except where annual stress test demonstrates absence of structural flaws. Note: Rotor life span may be reduced or warranty voided if autoclaved; contact manufacturer for guidance.

2. ''Planning for Use''

A. Complete lab-specific training for the centrifuge.

B. Wear appropriate PPE: Including safety eyewear, gloves, lab coat, and appropriate street clothing (i.e., long pants and closed-toe shoes). Ensure gloves are compatible with hazard(s).

C. Inspect centrifuge:
* Ensure tubes are rated for intended use (speed, temperature, and chemical resistance).
* Ensure rotor is compatible with centrifuge and seated on the drive correctly.
* Ensure rotor and safety cups/buckets are free of cracks and deformities.
* Ensure rotor O-ring is not cracked, missing, or worn.
* Ensure safety cups/buckets are attached correctly and able to move freely.

Contact a qualified service technician if inspection identifies centrifuge components requiring repair or replacement.

D. Prepare centrifuge tubes for loading:
* Inspect centrifuge tubes before use.
* Follow manufacturer’s filling limits for tubes. Do not overfill or underfill tubes.
* For biohazardous materials, disinfect outside of tubes prior to removal from biosafety cabinet and loading into rotor.

When centrifuging hazardous materials, use tightly capped tubes and/or sealable safety cups or rotors that can be loaded and unloaded in a fume hood or biosafety cabinet, depending on hazard. If a specific procedure does not allow for this, contact Occupational Safety and Environmental Health (OSEH) at (734) 647-1143 for guidance.

E. Use in-line filter: For high speed centrifuges and ultracentrifuges, use in-line filters to prevent contamination of vacuum pump and pump oil. Provide secondary containment for vacuum pump.

3. ''Centrifuge Operation''

A. Balance centrifuge: Follow manufacturer’s instructions for proper centrifuge balancing steps.

B. Start run
* Do not leave centrifuge until full operating speed is reached and it appears to be running safely without incident.
* Stop centrifuge immediately if you notice any unusual noises or shaking. Confirm rotor is balanced.

To prevent rotor failure, do not exceed maximum speed and maximum mass limits for the rotor. You must reduce rotor speed if sample density calculations indicate maximum mass limits will be exceeded; contact manufacturer for guidance.

4 ''Sample Removal''

A. Stop run: Ensure centrifuge comes to complete stop before opening cover. When centrifuging hazardous materials, wait at least 10 minutes after run to allow aerosols to settle before opening centrifuge.

B. Check for leaks/spills: In samples, rotor, safety cups/buckets, and centrifuge well.

C. Open sealable tubes/safety cups/rotors: Wear appropriate PPE and open inside fume hood or biosafety cabinet, depending on hazard.

==Personal Protective Equipment==
When operating the centrifuge, proper eye protection (glasses/goggles), gloves, lab coat, and close-toe footwear should be worn. Gloves should be appropriate for the present hazard.

==Waste Disposal==
Centrifuge Disposal

1. For biohazardous materials, clean/disinfect centrifuge and remove/cross out biohazard sticker. Attach note on centrifuge describing what has been done.

2. For radioactive materials, request radiation safety survey and signage before disposal of centrifuge.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

TREATMENT FACILITIES:
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for more information.

==Spill Procedure==
1. Mechanical Failure

Turn off centrifuge immediately and unplug power cord. Do not use centrifuge again until inspected by qualified service technician.

2. Hazardous Material Spill/Exposure

A. Turn off centrifuge immediately. Keep centrifuge cover closed for at least 30 minutes to reduce aerosolization of hazardous material.

B. Follow spill, exposure, and incident reporting instructions.
* For chemical, radioactive, and biohazard spills, contact OSEH at (734) 647-1143 for assistance and guidance.

Remember:
* Conduct rotor or safety cup/bucket cleanup in nearest biosafety cabinet or fume hood, depending on hazard.
* Use tongs or forceps to avoid contact with sharp-edged debris.
* Avoid alkaline cleaners for aluminum centrifuge components.
* Avoid abrasive wire brushes for cleaning.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling centrifuges.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Ccousz|Ccousz]] ([[User talk:Ccousz|talk]]) 17:19, 14 October 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:18, 10 November 2016 (UTC)
*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:09, 5 June 2017 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 18:49, 7 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:01, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:59, 12 June 2017 (UTC)

Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☒ NO

Laboratory Director - Dave Bridges Revision Date - 2016-10-14

SOP - Electrophoresis

2017-06-12T21:57:19Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
This standard operating procedure outlines the process of electrophoresis, including the use of equipment and chemicals necessary for the process. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when performing electrophoresis.

Electrophoresis uses electrical energy to separate molecules based on their size, structure, and electrical charge.

==Potential Hazards==
Electrophoresis equipment can pose significant electrical hazard in the laboratory. Typical electrophoresis units operating at 100 volts can provide a lethal shock of 25 milliamps. Take the following precautions:

'''Power Supplies:'''
*Inspect to ensure all switches and indicators are in proper working condition and that power cords and leads are undamaged and properly insulated.
*Label equipment with warning: “Danger Electrical Hazard.”
*Connect to ground fault circuit interrupters (GFCIs).
*Use 3-prong plugs.
*Use power supplies with safety features that detect no-load, overload, sudden load change, short circuit, arc or ground leak, etc.

'''Connecting Leads:'''
*Turn off main power supply before connecting or disconnecting electrical leads.
*With dry gloved hands, connect one lead at a time using one hand only.
*Be sure that leads/banana plugs are fully seated.

'''Using Equipment:'''
*Don’t run equipment unattended.
*Keep equipment clear of unintentional grounding points and conductors (e.g., sinks or other water sources, metal plates, jewelry, aluminum foil, pipes or other electrical/metal equipment).
*Gel chamber must have a lid or cover with safety interlocks to prevent accidental contact with energized electrodes or buffer solutions.
*Gel chamber exterior must be dry with no spilled solutions. Check for leaks.
*Switch off all power and unplug the leads before opening the gel chamber lid or reaching inside the gel chamber.
�
'''Hazardous chemicals'''
Commonly used in conjunction with electrophoresis work include:
*Ethidium bromide – mutagen, irritant
*Acrylamide – carcinogen, neurotoxin, irritant
*Phenol – corrosive, toxic
*Chloroform – suspect carcinogen, toxic

Always review the Safety Data Sheet prior to working with any hazardous material.
Laboratory personnel may be exposed to thermal hazards posed by liquefied gels.

'''Ultraviolet''' (UV) light boxes are often used in visualizing ethidium bromide gels and pose potential exposures to UV radiation.

==Engineering and Work Practice Controls==
*Read and follow manufacturer’s instructions for electrophoresis equipment.
*Prepare Standard Operating Procedure relevant to health and safety.
*Consult with Lab Director prior to initial use of electrophoresis equipment. Discussion should include special hazards and safety precautions.
*Measure, mix and handle all hazardous powdered chemicals or gel prep mixtures with hazardous components (e.g., acrylamide monomer, ethidium bromide, phenol, ammonium persulfate, and formaldehyde) in a fume hood.
*Purchase pre-made gels or pre-mixed acrylamide and ethidium bromide solutions instead of making your own.
*Consider using ethidium bromide substitutes.
*Exercise caution when using microwave to liquefy gels – don’t use sealed containers, beware of superheated liquids that may froth up unexpectedly. Let hot gel preps cool to 50°-60°C before adding ethidium bromide or pouring into trays. Wear insulated gloves and point the flask opening away from you.

==Protective Equipment==
*Wear lab coat with fully extended sleeves, safety glasses or splash goggles if appropriate, nitrile gloves (latex is not effective), pants, and closed-toe shoes.
*Wear appropriate skin and eye protection for UV radiation work.

==Waste Disposal==
'''Hazardous Waste Management:''' Handle and store hazardous waste following the guidelines above for work practice controls, transportation and storage. Contact OSEH Hazardous Materials Management (HMM) at 734-763-4568 with questions and to schedule a pickup of hazardous waste. For more information regarding chemical waste, refer to the HMM section on OSEH’s website at http://www.oseh.umich.edu/hazmats/chemical.shtml.

'''Non-Hazardous Waste Management:''' Some gels may be considered non-hazardous but still present a health hazard if untreated or discarded into the normal trash. Contact HMM for proper collection and disposal.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.

For an actual chemical exposure/injury:
*Remove contaminated clothing. Flush exposed eyes or skin with water for at least 15 minutes. Seek medical attention (see below).
*For inhalation exposure, remove all persons from the contaminated area. Get medical aid.
*If an ambulance is needed, call the University of Michigan Division of Public Safety and Security (DPSS) at 911 to request assistance.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==Treatment Facilities:==
*U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
*University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
*UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

Click here for more information.

==Spill Procedure==
*When a spill occurs, personal safety should always come first.
*Alert and clear everyone in the immediate area where the spill occurred.
*Follow spill procedures described in the SOP and SDS for the particular chemical spilled.

'''Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.
'''

==Training of Personnel==

All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when performing electrophoresis.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 15:00, 13 October 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:19, 10 November 2016 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 16:15, 31 October 2016 (UTC)
*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:10, 5 June 2017 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:12, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:57, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☒ NO

Laboratory Director - Dave Bridges Revision Date - 2016-10-13

SOP - Flammables and Combustibles

2017-06-12T21:54:49Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
This standard operating procedure outlines the handling and use of flammable and combustible materials. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling flammable and combustible materials.

Flammable and combustible liquids can burn and are classified, or grouped, as either flammable or combustible by their flashpoints. Flammable liquids will ignite and burn easily at normal working temperatures. Combustible liquids have the ability to burn at temperatures that are usually above working temperatures. Flammable liquids have a flashpoint below 37.8°C (100°F) while Combustible liquids have a flashpoint at or above 37.8°C (100°F) and below 93.3°C (200°F).

==Potential Hazards==
Flammability - Check Safety Data Sheet of the material for other hazards.
Definitions –
* Flammable gas – a gas that, at 68°F or less and standard pressure, forms a flammable mixture with air at a concentration of 13% by volume or less OR that, at ambient temperature and pressure, forms a range of flammable mixtures with air wider than 12% by volume, regardless of the lower limit.
* Flammable liquid – a liquid having a flash point below 100°F.
* Flammable solid – a solid, other than a blasting agent or explosive, that (1) has an ignition temperature below 212°F, or (2) is capable of causing a fire through friction, absorption of moisture, or spontaneous chemical change, or (3) burns so vigorously and persistently as to create a serious hazard.
* Combustible liquid – a liquid having a flash point at or above 100°F.

==Engineering Controls==
Work in a chemical fume hood if air concentrations above 10% of the lower flammable limit may be reached, if the chemical is irritating to the eyes and respiratory system, and/or is toxic by inhalation.

==Work Practice Controls==
* Alert others in your lab of the materials you will be working with.
* Review SDSs before working with material.
* Know the location of the nearest fire extinguisher before beginning work.
* Use in the smallest practical quantities for the experiment being performed.
* Avoid using ignition sources (Bunsen burners, hot plates, electrical equipment with frayed or cracked wiring, etc.) and/or creating static electricity in areas where highly flammable chemicals are used.
* Ensure proper grounding. Be sure to ground metal containers when transferring flammable liquids.
* Keep containers of flammable chemicals closed at all times when not in use to prevent accumulation of flammable vapor concentrations.

==Personal Protective Equipment==
* Wear nitrile laboratory gloves, lab coat (flame-resistant lab coats for handling large volumes), safety glasses, clothing that minimizes exposed skin, and close-toed shoes for all work with flammables. If working with flammable liquids that may splash, wear chemical splash goggles.
* If a flammable liquid is also toxic, check glove guide for appropriate gloves and/or minimize glove contact with the liquid.
* Avoid wearing flammable clothing (such as many synthetics) when working with flammable materials.

Gloving Material Advantages and Disadvantages
BUTYL RUBBER
* Good for ketones and esters
* Poor for gasoline and aliphatic, aromatic, and halogenated hydrocarbons
* Expensive
* Only available as re-useable
* Poor touch sensitivity
NEOPRENE
* Good for acids, bases, alcohols, fuels, peroxides, hydrocarbons, and phenols
* Poor for halogenated and aromatic hydrocarbons
VITON
* Good for chlorinated and aromatic solvents
* Low surface tension repels most liquids
* Good resistance to cuts and abrasions
* Poor for ketones
* Expensive
* Only available as re-useable
* Poor touch sensitivity
VINYL - Polyvinyl Alcohol (PVA)
* Good for aromatic and chlorinated solvents
* Poor for water-based solutions (dissolves in water)
* Avoid: Contact with water or water-based solutions, water solubles
* Only available as re-useable
* Poor touch sensitivity

==Transportation and Storage==
* Flammable chemicals should be stored in appropriate areas within the laboratory and away from any potentially incompatible materials.
* Transport flammables in secondary containment, preferably a polyethylene or solvent bottle carrier.
* Suitable fire control devices (such as fire extinguishers) must be available at locations where flammable or combustible liquids are stored.
* Open flames shall not be permitted in flammable liquid storage areas. Flammable and combustible liquids must not be stored in a manner that hinders safe egress.
* If large quantities (>60 gallons) of flammable or combustible liquids are stored, a specially-designed storage room may be required.
* Store these materials in a separate room from water-reactives.
* Flammable and combustible liquids may be stored in a flammable storage cabinet. All such storage cabinets must meet NFPA 30 requirements.
* Flammable liquids shall not be stored in unapproved or residential-type refrigerators.
* Secondary containment is recommended for liquids.
* Avoid storing on the floor.

==Waste Disposal==
Because most spent, unused and expired chemicals/materials are considered hazardous wastes, they must be properly disposed of. Do not dispose of chemical wastes by dumping them down a sink, flushing in a toilet or discarding in regular trash containers, unless authorized by OSEH Hazardous Materials Management (HMM). Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.

* If skin contact occurs, immediately remove contaminated clothing and rinse with water for at least 15 minutes.
* For eye exposures, immediately rinse eyes with copious amounts of water for at least 15 minutes, while occasionally lifting upper and lower lids, then promptly seek medical attention.
* If large amounts of vapors are inhaled, move person to fresh air immediately and seek medical attention.
* If ingested, seek medical attention immediately.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

TREATMENT FACILITIES:
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for additional accident and injury reporting information.

==Spill Procedure==
When a spill occurs, personal safety should always come first. Alert and clear everyone in the immediate area where the spill occurred.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., less than 1 gallon or 3.5 liters. A major/large chemical spill requires active assistance from emergency personnel.

Spill Response Steps:

MINOR CHEMICAL SPILL
* Alert people in immediate area of spill.
* Turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
* Open outside windows, if possible.
* Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.
* Avoid breathing vapors from spill.
* Confine spill to as small an area as possible.
* Do not wash spill down the drain.
* Use appropriate spill kits/sorbents to neutralize corrosives and/or absorb spill. Collect contaminated materials and residues and place in container and contact OSEH-HMM (734) 763-4568 for proper disposal.
* Clean spill area with water.

MAJOR CHEMICAL SPILL
* Attend to injured or contaminated persons and remove them from exposure.
* Alert people in the laboratory to evacuate.
* If appropriate turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
* Call University of Michigan Police Department (UMPD) at 911 immediately for assistance.
* Close doors to affected area.
* Post warnings to keep people from entering the area.
* Have person available that has knowledge of incident and laboratory to assist emergency personnel.

==Additional Spill Information Links:==
* www.oseh.umich.edu/pdf/chemspil.pdf
* http://www.oseh.umich.edu/emer-chemical.shtml.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Ccousz|Ccousz]] ([[User talk:Ccousz|talk]]) 17:09, 12 October 2016 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 16:17, 31 October 2016 (UTC)
* [[User:Davebridges|Davebridges]] ([[User talk:Davebridges|talk]]) 14:51, 8 November 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:18, 10 November 2016 (UTC)
*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:11, 5 June 2017 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:15, 7 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:03, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:54, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO
Laboratory Director - Dave Bridges Revision Date - 2016-10-12

SOP - Rapamycin

2017-06-12T21:53:33Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
''This standard operating procedure outlines the handling and use of rapamycin. Review this document and supply the information required in order to make it specific to your facility. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling rapamycin.''

Rapamycin is an immunosuppressant drug used to prevent rejection in organ transplantation; it is especially useful in kidney transplants. Rapamycin is not a calcineurin inhibitor. However, it has a similar suppressive effect on the immune system. Rapamycin inhibits the response to interleukin-2 (IL-2) and thereby blocks activation of T- and B-cells. The health effects from long-term usage could include increase the risk of developing an infection or cancer, especially lymphoma (cancer of a part of the immune system) or skin cancer.

Synonyms: RAPA, RPM, Rapamune, Sirolimus, "macrocyclic polyketide", "macrocyclic triene antibiotic", antirestenotic, immunosuppressant, "mTOR inhibitor"

==Potential Hazards==
*Toxic: danger of serious damage to health by prolonged exposure if swallowed.
*Possible carcinogenic effect.
*Biologically active compounds may produce idiosyncratic effects which are entirely unpredictable on the basis of literature searches and prior clinical experience (both recent and past).
*Ingestion of the material may be damaging to the health of the individual.

Macrolides comprise a large group of antibiotics derived from Streptomyces spp. having in common a macrocyclic lactone ring to which one or more sugars are attached. They are all weak bases. The most common side effect produced by the family of macrolide antibiotics is gastrointestinal discomfort. Supra-infections may occur although these are rare. Several macrolides produced allergic sensitization but, again, these are rare. Symptoms include watery eyes, shortness of breath, nasal congestion, choking, coughing and wheezing. Allergic skin reactions have also occurred. Exposure to at least one member of the family, erythromycin, at high concentrations, has produced reversible deafness (ototoxicity). Systemic reactions including fever, rash, and lymph-node pain or swelling have been produced by the avermectin group. Ivermectin has produced ataxia (incoordination), lethargy, bradypnea (slowed breathing), vomiting, mydriasis (dilated pupils), sedation, tremors and death in animals. The avermectin group (anthelmintics, insecticides and acaricides) mediate the transmission of gamma-butyric acid (GABA), an inhibitory neurotransmitter, in mammals thus causing paralysis. Hepatotoxic effects with transient disturbances and jaundice have resulted from the use of oleandomycin. Transient alterations in heart rate/ rhythm have also been produced by several members of the family (notably tilmicosin). Heart muscle degeneration, characterized by small areas of cell death has also been reported in animals exposed to tilmicosin. Cross-resistance is often observed between the macrolide, lincosamide and streptogramin group of antibiotics.

'''Eye:'''
Although the material is not thought to be an irritant, direct contact with the eye may cause transient discomfort characterized by tearing or conjunctival redness (as with windburn). Slight abrasive damage may also result. The material may produce foreign body irritation in certain individuals.

'''Skin:'''
The material is not thought to produce adverse health effects or skin irritation following contact (as classified using animal models). Nevertheless, good hygiene practice requires that exposure be kept to a minimum and that suitable gloves be used in an occupational setting.

Open cuts, abraded or irritated skin should not be exposed to this material. Entry into the blood-stream through, for example, cuts, abrasions or lesions, may produce systemic injury with harmful effects. Examine the skin prior to the use of the material and ensure that any external damage is suitably protected.

'''Inhaled:'''
The material is not thought to produce either adverse health effects or irritation of the respiratory tract following inhalation (as classified using animal models). Nevertheless, adverse effects have been produced following exposure of animals by at least one other route and good hygiene practice requires that exposure be kept to a minimum and that suitable control measures be used in an occupational setting.

Persons with impaired respiratory function, airway diseases and conditions such as emphysema or chronic bronchitis, may incur further disability if excessive concentrations of particulate are inhaled.

==Engineering Controls==
*Enclosed local exhaust ventilation is required at points of dust, fume or vapor generation.
*Work should be performed in a fume hood or Biological Safety Cabinet.

==Work Practice Controls==
Decontamination should consist of surface cleaning with water and detergent followed by thorough rinsing. The use of detergent is recommended because there is no single accepted method of chemical deactivation for all agents involved. A plastic backed absorbent pad should be placed under the work area during the process. This should be changed at the end of each procedure or when a spill occurs.

==Personal Protective Equipment (PPE)==
Personal protective equipment includes a lab coat, eye protection and disposable gloves. Gloves should be changed frequently and should be pulled up over sleeves if possible to reduce the likelihood of any exposed skin. Wash hands and arms immediately after working with the substance. Particulate respirator protection should be assessed where incidental or accidental exposure is anticipated.

Note: Respirators are masks designed to protect the wearer from specific airborne hazards and are different from surgical masks, which protect the wearer only from splashes and are primarily intended to protect others from infectious aerosols exhaled by the wearer. Respirator use requires employee participation in the Respiratory Protection Program, which involves medical clearance and annual fit testing and training. Please be clear about use of surgical masks versus respirators. (Do NOT use the vague term “masks”.)

==Transportation and Storage==
*Keep containers tightly closed and store in a dry and ventilated area.
*Store away from strong oxidizing agents.
*Store according with information on product insert.
*Rapamycin is NOT REGULATED by DOT or IATA.

==Waste Disposal==
All hazardous chemical agent contaminated waste should be placed in a 5 gallon white pail. The container should be closed except when actively adding waste. The container must be located in the area where hazardous drugs are being used. All items contaminated by hazardous drugs, including gloves, syringes, vials needles, and solution containers should be disposed according to OSEH guidelines.

Sharps – place needles, syringes with needles attached and other breakable items into appropriately labeled sharps containers.

Empty stock vials, reagent bottles, etc. – triple rinse with copious amounts of water. Deface label with black magic marker or scraper. Place in a cardboard box for disposal.

Because most spent, unused and expired chemicals/materials are considered hazardous wastes, they must be properly disposed of. ''Do not dispose of chemical wastes by dumping them down a sink, flushing in a toilet or discarding in regular trash containers, unless authorized by OSEH Hazardous Materials Management (HMM).'''''Bold text''' Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

If animals are administered rapamycin in their food or water, both the food and water container must be labeled. Food or water that is not consumed by the animals must be collected, labeled and submitted for disposal as a hazardous material.

==Exposures/Unintended Contact==

'''If the employee is in need of emergency medical attention, call 911 immediately.
'''

'''SWALLOWED'''
*Seek medical advice.
*Give water to rinse out mouth.
*If swallowed do NOT induce vomiting, unless directed to do so by medical personnel.
*If vomiting occurs, lean patient forward or place on left side (head-down position, if possible) to maintain open airway and prevent aspiration.

'''EYE'''

If this product comes in contact with the eyes:
*Wash out immediately with fresh running water.
*Ensure complete irrigation of the eye by keeping eyelids apart and away from eye and moving the eyelids by occasionally lifting the upper and lower lids.
*If pain persists or recurs seek medical attention.
*Removal of contact lenses after an eye injury should only be undertaken by skilled personnel.

'''SKIN'''

If skin or hair contact occurs:
*Flush skin and hair with running water (and soap if available).
*Seek medical attention in event of irritation.

'''INHALED'''
*If inhaled remove from contaminated area.
*Consult a physician.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==Treatment Facilities:==
*U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
*University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
*UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

Click here for additional accident and injury reporting information.

==Spill Procedure==
• When a spill occurs, '''personal safety should always come first'''''Italic text''.
• Alert and clear everyone in the immediate area where the spill occurred.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel. A '''major/large chemical spill'''''Italic text'' requires active assistance from emergency personnel.

==Spill Response Steps:==

'''MINOR CHEMICAL SPILL'''
*Alert people in immediate area of spill.
*Open outside windows, if possible.
*Use proper personal protective equipment (PPE) as indicated above.
*Avoid breathing vapors from spill.
*Confine spill to as small an area as possible.
*Do not wash spill down the drain.
*Use appropriate spill kits/sorbents to absorb spill. Collect contaminated materials and residues and place in container. For powdered chemicals sweep carefully to avoid generation of dust or, if appropriate, use moist sorbent pads or wet the powder with a suitable solvent and then wipe with a dry cloth. Contact OSEH-HMM (734) 763-4568 for proper disposal.
*Clean spill area with water.

'''MAJOR CHEMICAL SPILL'''
*Attend to injured or contaminated persons and remove them from exposure.
*Alert people in the laboratory to evacuate.
*If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
*Call University of Michigan Division of Public Safety and Security (DPSS) at 911 immediately for assistance.
*Close doors to affected area.
*Post warnings to keep people from entering the area.
*Have person available that has knowledge of incident and laboratory to assist emergency personnel.

==Additional Spill Links:==
*www.oseh.umich.edu/pdf/chemspil.pdf
*http://www.oseh.umich.edu/emer-chemical.shtml.

'''Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.
'''

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling rapamycin.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 16:12, 13 October 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:37, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:53, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☒ NO

Laboratory Director - Dave Bridges Revision Date - 2016-10-13

SOP - Carcinogens

2017-06-12T21:51:57Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

'''Laboratory Director (LD) Approval is Required Prior to Performing this Procedure
'''
==Description==
This standard operating procedure is designed to provide guidance in writing procedures for the safe handling and disposal of carcinogens, which are regarded as particularly hazardous substances. Review this document and supply the information required in order to make it specific to your laboratory.

A carcinogen is defined by meeting one the following:
*It is listed under the category, “known to be carcinogens” in the latest National Toxicology Program annual report.
*It is listed under Group 1 by the International Agency for Research on Cancer (IARC).
*It is listed in Group 2A or 2B by IARC or under the category, “reasonably anticipated to be carcinogens” by NTP, and causes statistically significant tumor incidence in experimental animals in accordance with any of the following criteria:
**After inhalation exposure of 6-7 hours per day, 5 days per week, for a significant portion of a lifetime to dosages of less than 10 mg/m3.
**After repeated skin application of less than 300 mg/kg of body weight per week.
**After oral dosages of less than 50 mg/kg of body weight per day.
If you have questions concerning the applicability of any item listed in this procedure contact the LD/Laboratory Supervisor or Occupational Safety and Environmental Health at (734) 647-1143.

==Potential Hazards==
A carcinogen commonly describes any agent that can initiate or speed the development of malignant or potentially malignant tumors, malignant neoplastic proliferation of cells, or cells that possess such material.

==Engineering Controls==
*Manipulation of carcinogens should be carried out in a fume hood. If the use of a fume hood proves impractical refer to the section on special ventilation.
*Certain carcinogens must be handled in a glove box rather than a fume hood. Occupational Safety and Environmental Health (734) 647-1143 or the LD will determine if this is required.
*Where the eyes or body of any person may be exposed to carcinogens, suitable facilities for quick drenching or flushing of the eyes and body shall be provided within the work area for immediate emergency use. Bottle type eyewash stations are not acceptable.
*Safety shielding is required any time there is a risk of explosion, splash hazard or a highly exothermic reaction. All manipulations of carcinogens which pose this risk should occur in a fume hood with the sash in the lowest feasible position. Portable shields, which provide protection to all laboratory occupants, are acceptable.
*Manipulation of carcinogens outside of a fume hood may require special ventilation controls in order to minimize exposure to the material. Fume hoods provide the best protection against exposure to carcinogens in the laboratory and are the preferred ventilation control device. When possible, handle carcinogens in a fume hood. If the use of a fume hood proves impractical, attempt to work in a glove box or on an isolated area on the bench top.
*If available, consider using a Biological Safety Cabinet. The biological safety cabinet is designed to remove particulates (the carcinogen) before the air is discharged into the environment. Carcinogens that are volatile must not be used in a biological safety cabinet unless the cabinet is vented to the outdoors.
*If your research does not permit the handling of carcinogens in a fume hood, biological safety cabinet, or glove box, you must contact Occupational Safety and Environmental Health (OSEH).
*Evacuated glassware can implode and eject flying glass, and splattered chemicals. Vacuum work involving carcinogens must be conducted in a fume hood, glove box or isolated in an acceptable manner.
*Mechanical vacuum pumps must be protected using cold traps and, where appropriate, filtered to prevent particulate release. The exhaust for the pumps must be vented into an exhaust hood.

==Work Practice Controls==
All areas where carcinogens are stored or manipulated must be labeled as a designated area.
*The room sign for the laboratory must contain a “Designated Area Within” identifier.
*All locations within the laboratory where carcinogens are handled should be demarcated with designated area caution tape (available from OSEH, or your department) and/or posted with designated area caution signs. This includes all fume hoods and bench tops where the carcinogens are handled.
*All employees with access to designated areas must be made aware of the substances being used and necessary precautions by the LD.
*Where feasible, carcinogens should be manipulated over plastic-backed disposable paper work surfaces. These disposable work surfaces minimize work area contamination and simplify clean up.

'''Doorways:''' The room sign must contain a “Designated Area Within, Caution” where carcinogens, reproductive hazards, and/or acutely toxic chemicals are stored or used.

'''Containers:''' All containers of carcinogens must be clearly labeled with the correct chemical name. Handwritten labels are acceptable; chemical formulas and structural formulas are not acceptable.

Wash hands, forearms, face, and neck upon exit of a designated area to decontaminate.

Keep accurate records of amounts of carcinogenic substances being stored and used, including dates of use and names of researchers using the substances.

==Personal Protective Equipment==
Eye protection in the form of safety glasses must be worn at all times when handling carcinogens. Ordinary (street) prescription glasses do not provide adequate protection. (Contrary to popular opinion these glasses cannot pass the rigorous test for industrial safety glasses.) Adequate safety glasses must meet the requirements of the Practice for Occupational and Educational Eye and Face Protection (ANSI Z.87. 1) and must be equipped with side shields. Safety glasses with side shields do not provide adequate protection from splashes; therefore, when the potential for splash hazard exists, other eye protection and/or face protection must be worn.
Gloves must be worn when handling carcinogens. Disposable nitrile gloves provide adequate protection against accidental hand contact with small quantities of most laboratory chemicals. Lab workers should contact OSEH for advice on chemical resistant glove selection when direct or prolonged contact with hazardous chemicals is anticipated.
Lab coats, closed toed shoes and long sleeved clothing must be worn when handling carcinogens. Additional protective clothing should be worn if the possibility of skin contact is likely.

==Waste Disposal==
Because most spent, unused and expired chemicals/materials are considered hazardous wastes, they must be properly disposed of. Do not dispose of chemical wastes by dumping them down a sink, flushing in a toilet or discarding in regular trash containers, unless authorized by OSEH Hazardous Materials Management (HMM). Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

'''If the employee is in need of emergency medical attention, call 911 immediately.
'''

For an actual chemical exposure/injury:
*Remove contaminated clothing. Flush exposed eyes or skin with water for at least 15 minutes. Seek medical attention (see below).
*For situations with risk of inhalation exposure (including spills of powder outside of a chemical fume hood), remove all persons from the contaminated area.
*If an ambulance is needed, call the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 to request assistance.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==Treatment Facilities:==
*U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
*University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
*UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

Click here for more information.

==Spill Procedures==
*When a spill occurs, personal safety should always come first.
*Alert and clear everyone in the immediate area where the spill occurred.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., less than 1 liter. A major/large chemical spill requires active assistance from emergency personnel.

==Additional Spill Response Steps:==

'''MINOR CHEMICAL SPILL'''
*Alert people in immediate area of spill.
*If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
*Open outside windows, if possible.
*Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.
*Avoid breathing vapors from spill.
*Confine spill to as small an area as possible.
*Do not wash spill down the drain.
*Use appropriate spill kits/sorbents to neutralize corrosives and/or absorb spill. Collect contaminated materials and residues and place in container. For powdered chemicals sweep carefully to avoid generation of dust or, if appropriate, use moist sorbent pads or wet the powder with a suitable solvent and then wipe with a dry cloth. Contact OSEH-HMM (734) 763-4568 for proper disposal.
*Clean spill area with water.

'''MAJOR CHEMICAL SPILL'''
*Attend to injured or contaminated persons and remove them from exposure.
*Alert people in the laboratory to evacuate.
*If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
*Call University of Michigan Division of Public Safety and Security (DPSS) at 911 immediately for assistance.
*Close doors to affected area.
*Post warnings to keep people from entering the area.
*Have person available that has knowledge of incident and laboratory to assist emergency personnel.

'''Additional Spill Links:'''
*www.oseh.umich.edu/pdf/chemspil.pdf
*http://www.oseh.umich.edu/emer-chemical.shtml.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling carcinogens.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 13:32, 13 October 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:18, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 18:48, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:51, 12 June 2017 (UTC)
Laboratory Director - Dave Bridges Revision Date - 2016-10-13

SOP - Formalin and Paraformaldehyde

2017-06-12T21:50:59Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

'''Laboratory Director (LD) Approval is Required Prior to Performing this Procedure'''

==Description==
This standard operating procedure outlines the handling and use of formalin and paraformaldehyde. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling formalin and paraformaldehyde.

Formalin and paraformaldehyde are primarily used as fixatives. These fixatives act to preserve and stabilize cells and tissues prior to examination processes. The aqueous solution of formalin is 37-40 percent formaldehyde in water or methanol. Paraformaldehyde is the crystallized polymer of formaldehyde (97%) that is weighed out and dissolved in solution for experimentation or for cell and tissue fixation. Typically 3-10% formalin or paraformaldehyde solutions are used to perfuse or fix tissues.

==Useful Formalin and Paraformaldehyde Links:==
http://www.cdc.gov/niosh/docs/81-111/

http://www.cdc.gov/niosh/topics/formaldehyde/

http://www.cdc.gov/niosh/npg/npgd0293.html

==Potential Hazards==
*Formalin and paraformaldehyde solutions can emit formaldehyde gas, a known human carcinogen, and can irritate the eyes and skin.
*Working with paraformaldehyde powder (and, to a lesser extent, flakes or granules), can expose employees to paraformaldehyde dust, which is a strong irritant/sensitizer.
*Contact with these solutions or paraformaldehyde solids may also cause drying of the skin and/or allergic dermatitis.
*The MIOSHA Permissible Exposure Limit for formaldehyde is 0.75 ppm for 8 hours or 2 ppm for 15 minutes. There is a substance-specific MIOSHA standard for formaldehyde, and an action limit of 0.5 ppm.
*Consult your Safety Data Sheet for more information on hazards.

==Engineering Controls==
*Work with concentrated (>4% formaldehyde/paraformaldehyde) solutions only in a chemical fume hood.
*Handle paraformaldehyde powder (and, preferably, granules or flakes) only in a chemical fume hood.
*Dilute solutions (<4% formaldehyde) may be used on the benchtop in small quantities.
*If there is any possibility that an employee's eyes may be splashed with solutions containing 0.1 percent or greater formaldehyde, an eyewash/drench hose must be available within the immediate work area for emergency use.
*If employees' skin may become splashed with solutions containing 1 percent or greater formaldehyde, for example, because of equipment failure or improper work practices, the MIOSHA formaldehyde standard requires a conveniently-located safety shower. Contact Occupational Safety and Environmental Health (OSEH) at (734) 647-1143 to determine if a safety shower will be needed.

==Work Practice Controls==
*Designate an area for working with concentrated formalin, concentrated paraformaldehyde solutions, and paraformaldehyde solid, and label it as such.
*Keep containers closed as much as possible.
*Use in the smallest practical quantities for the experiment being performed.
*If you are weighing paraformaldehyde powder and the balance cannot be located in a fume hood or BSC, tare a container then add powder in the hood and cover before returning to the balance to weigh the powder.
*Labs handling moderate to large quantities of formaldehyde-containing solutions on a regular basis should contact OSEH at (734) 647-1143 for assessment of exposure. Areas that handle only small (100 ml or less) pre-filled specimen containers, or that work with formaldehyde-containing solutions exclusively in a functioning chemical fume hood, would have low potential for overexposure, but should contact OSEH if there are concerns.
*Once work with formalin/paraformaldehyde is complete, wipe down area with a soap and water solution.

==Protective Equipment==
Wear standard nitrile laboratory gloves, chemical splash goggles, face shield, and lab coat. If splash may occur, also wear an impervious apron. (MIOSHA requires that all contact of the eyes and skin with liquids containing 1 percent or more formaldehyde be prevented by the use of chemical protective clothing made of material impervious to formaldehyde and the use of other personal protective equipment, such as goggles and face shields, as appropriate to the operation.)

==Transportation and Storage==
*Transport formaldehyde solutions in secondary containment, preferably a polyethylene or other non-reactive acid/solvent bottle carrier.
*Keep container in cool, well-ventilated area.
*Keep container tightly closed and sealed until ready for use.
*Store in secondary containment with flammables, away from oxidizers, reducing agents, metals, and acids.
*Keep containers of paraformaldehyde (PFA) solid away from water.
*Avoid storing on the floor.
*Avoid ignition sources.

==Waste Disposal==
Formalin and paraformaldehyde solutions and powders must be disposed following the guidelines above while accumulating wastes and awaiting chemical waste pickup. Waste must be disposed of following OSEH Hazardous Materials procedures. Contact OSEH’s Hazardous Materials Management (HMM) at (734) 763-4568 for waste containers, labels, manifests, and waste collection. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

'''If the employee is in need of emergency medical attention, call 911 immediately.
'''
For an actual chemical exposure/injury,
*Flush exposed eyes or skin with water for at least 15 minutes, then seek medical attention (see below).
*If there is respiratory irritation associated with exposure, remove all persons from the contaminated area.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==Treatment Facilities:==
*U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
*University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
*UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

Click here for more information.

==Spill Procedure==
Employees in the area should be prepared to clean up minor spills, including most spills confined to the chemical fume hood. Wearing double nitrile gloves, splash goggles, face shield and lab coat (and impermeable apron, if available), use absorbent pads to absorb spilled material. For small spills of solid PFA, dampen the absorbent pad with methanol before placing over the spilled material and allow to sit for a few minutes before wiping up. After spill has been completely absorbed, wash down contaminated area with soap and water at least two times. Contaminated PPE and clean-up materials must be placed in a clear plastic bag or compatible container for pick-up by OSEH.

==Additional Spill Links:==
*www.oseh.umich.edu/pdf/chemspil.pdf
*http://www.oseh.umich.edu/emer-chemical.shtml.

For large spills and most spills of formalin or paraformaldehyde solutions or paraformaldehyde powder that occurs outside of a chemical fume hood should be referred to OSEH-HMM.

'''Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.
'''

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling formalin and paraformaldehyde.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 15:58, 13 October 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:23, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:31, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:50, 12 June 2017 (UTC)
Laboratory Director - Dave Bridges Revision Date - 2016-10-13

SOP - Ethidium Bromide

2017-06-12T21:49:29Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
This standard operating procedure outlines the handling and use of Ethidium Bromide (EtBr). Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling EtBr.

EtBr intercalates double-stranded DNA and RNA and acts as a frameshift mutagen. It can also be used in conjunction with acridine orange to differentiate between viable, apoptotic and necrotic cells.

==Safer Alternative to Ethidium Bromide==
A recommended, safer alternative to ethidium bromide is using Gel Red as a gel staining agent for nucleic acid work (dsDNA, ssDNA or RNA) in agarose gels or polyacrylamide gels. It is not only highly sensitive but also tested to be environmentally safe and non-hazardous. Click the following link below for more details: http://www.ocs.umich.edu/pdf/GelRedFactSheet.pdf

==Potential Hazards==
*EtBr is an odorless solid that is irritating to the eyes, skin, mucous membranes and upper respiratory tract.
*EtBr is a potent mutagen and should be treated as a possible reproductive hazard and carcinogen.
*Consult the SDS and Laboratory Chemical Safety Summary for Ethidium Bromide in Prudent Practices in the Laboratory (National Academies Press).

==Engineering Controls==
Stock solutions of EtBr should be prepared in a chemical fume hood. If aerosols may be produced (dust or liquid mist), EtBr must be handled in a chemical fume hood, exhausted biological safety cabinet with negative pressure ductwork, or other exhausted enclosure. Aerosols may be produced during any open handling of dry powder, and during open or pressurized manipulations of solutions.

==Work Practice Controls==
*Set up a designated area for work with Ethidium Bromide, and label it with the following wording: “Ethidium Bromide in use. Mutagen and irritant.”
*Line the work area with a disposable plastic-backed absorbent pad.
*Keep containers closed as much as possible.
*If weighing dry powders and the balance cannot be located in a fume hood or BSC, tare a container then add the material to the container in a hood and seal the container before returning to the balance to weigh the powder.
*Change gloves regularly (at least every two hours) and wash hands at the time of the glove change. No skin contact is permitted.

==Protective Equipment==
*Standard nitrile laboratory gloves and a fully buttoned lab coat with sleeves extending to the wrists should be worn when handling EtBr (powder and solutions).
*If splashes may occur, wear goggles and a face shield. Otherwise, wear standard laboratory safety glasses.
*When using a UV light to visualize EtBr contamination, wear UV-blocking eyewear or work behind a UV shielding glass. (Most standard safety glasses will block UV, but employees should check the approval of their safety glasses.)
*In cases where the arms or torso may be exposed to liquid suspensions or dry particles, wear Tyvek sleeves and/or gowns (or other air-tight non-woven textile).

==Transportation and Storage==
*EtBr powder and solutions should be in tightly closed, shatter-resistant containers during transportation and storage. Secondary containment is advised.
*It should be stored away from strong oxidizing agents in a cool, dry place.

==Waste Disposal==
EtBr waste in concentrated or solid form is collected as chemical waste and should not be flushed down the drain or disposed of in the trash. Go to Drain Disposal for more information. Waste should be properly labeled and handled.

'''Liquids:''' Non-aqueous solutions and solutions containing EtBr concentration will be picked up by OSEH Hazardous Materials Management (OSEH-HMM). OSEH-HMM provides 1-gallon or 5-gallon pails for solid waste and gels, 1-gallon glass jugs for liquid waste, and smaller waste containers. Go to Waste Supplies for more information.

'''Contaminated sharps (needles, syringes, slides, broken glass, etc.):''' Discard in an infectious waste sharps container clearly labeled "CHEMICAL CONTAMINATED SHARPS-DO NOT AUTOCLAVE". Contact OSEH-HMM for pickup of the sharps container as infectious waste without autoclaving within 90 days or when it is 2/3 to 3/4 full.

'''Solids (contaminated gloves, centrifuge tubes, towels, etc.):''' Store in a properly labeled 1-gallon or 5-gallon pail for disposal as chemical waste. Do not use glass containers.

'''Gels:''' Gels should be disposed of as contaminated solids described above.

Because most spent, unused and expired chemicals/materials are considered hazardous wastes, they must be properly disposed of. ''Do not dispose of chemical wastes by dumping them down a sink, flushing in a toilet or discarding in regular trash containers, unless authorized by OSEH Hazardous Materials Management (HMM).'''''Bold text''' Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended contact==

'''If the employee is in need of emergency medical attention, call 911 immediately.'''

'''For an actual chemical exposure/injury:'''
*Remove contaminated clothing. Flush exposed eyes or skin with water for at least 15 minutes. Seek medical attention (see below).
*For inhalation exposure, remove all persons from the contaminated area. Get medical aid.
*If an ambulance is needed, call the University of Michigan Division of Public Safety and Security (DPSS) at 911 to request assistance.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==TREATMENT FACILITIES:==
*U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
*University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
*UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for additional accident and injury reporting information.

==Spill Procedure==
*When a spill occurs, personal safety should always come first.
*Alert and clear everyone in the immediate area where the spill occurred.
*Small spills of EtBr solutions should be cleaned by laboratory staff. For large spills outside the fume hood, evacuate/restrict access to the laboratory and contact OSEH-HMM for assistance at (734) 763-4568.
*Individuals cleaning spills must wear appropriate protective equipment as described in the Personal Protective Equipment section of this document.
*Spills of EtBr solutions should be cleaned using absorbent pads followed by surface decontamination using soap and water. Spilled dry material should first be covered with moist absorbent pads to avoid generation of dust.
*Ensure all materials contaminated as a result of the cleanup process are collected and disposed of as hazardous waste as described in the EtBr Waste Disposal section of this document.

==Additional Spill Links:==
*www.oseh.umich.edu/pdf/chemspil.pdf
*http://www.oseh.umich.edu/emer-chemical.shtml.

'''Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.
'''

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling EtBr.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure.Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 15:48, 13 October 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:19, 10 November 2016 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 16:16, 31 October 2016 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:49, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☒ YES ☐ NO

SOP - BVT.2733

2017-06-12T21:48:12Z

Elhabbal:

OSEH Recommendations/Findings for BVT.2733

Potential hazards of this material are unknown. This may be due to lack of information provided on the specific drug and/or it may be investigational/experimental/novel in nature. Therefore, limited or no information regarding potential toxicity was found. Thus, it is always prudent to handle as potentially hazardous agents. Caution should be exercised when handling and preparing solutions. The lab must use a biological safety cabinet during reconstitution and dilution of this material as well as use proper techniques to prevent contamination onto work surfaces. Personnel preparing this agent should wear double nitrile or latex gloves, safety glasses and a lab coat. All needles, syringes, vials, ampules and other materials which have come in contact with this agent should be segregated and disposed as hazardous waste. Unless evidence is provided to support a different course of action, it is EHS’s policy to house animals in the ULAM Animal Containment Housing facilities for a period of 3 days after each administration. After this time, the animals must be changed to a clean cage and can then be moved to standard animal housing facility. All waste materials, including bedding and carcasses must be collected and disposed through EHS HazMat while the animals are in Containment Housing. EHS HazMat (3‐4568) must be contacted prior to the commencement of this experiment to ensure proper waste containers are available for collection of this material. This policy is designed to reduce the potential exposure of animal care technicians and cage wash personnel to hazardous materials that may contaminate the dirty bedding. EHS can reevaluate housing requirements if the lab can provide information indicating that the material(s) are metabolized to a non‐hazardous substance and poses little risk to animal care personnel. Follow the link below for help on the preparation of chemical waste for collection: http://www.ehs.umich.edu/pdf/Chemical_Waste_Poster.pdf

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:48, 12 June 2017 (UTC)

SOP - Bunsen Burners

2017-06-12T21:46:30Z

Elhabbal: /* Training of Personnel */

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
This standard operating procedure outlines the handling and use of Bunsen burners. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls and personal protective equipment when using Bunsen burners.

Bunsen burners produce a single open flame by burning a continuous stream of flammable gas used for heating, sterilization, and combustion.

==Potential Hazards==
Bunsen burners present burn and fire hazards due to the high-temperature open flame that is produced.

==Engineering Controls==
Biological safety cabinets (BSCs) and disposable sterile items obviate the need for open flames when aseptic conditions are needed. The use of a Bunsen burner in a BSC creates thermodynamic instabilities that interfere with the function of the unit, reducing its ability to provide the sterile field. If an open flame absolutely must be used in a BSC, recommended alternatives such as electrical incinerators or touch-plate microburners are available.

==Work Practice Controls==
Bunsen burner safety – best practice:
* PLACE the Bunsen burner away from any overhead shelving, equipment, or light fixtures.
* REMOVE all papers, notebooks, combustible materials and excess chemicals from the area.
* TIE-BACK any long hair, dangling jewelry, or loose clothing.
* INSPECT hose for cracks, holes, pinched points, or any other defect and ensure that the hose fits securely on the gas valve and the Bunsen burner.
* REPLACE all hoses found to have a defect before using.
* NOTIFY others in the laboratory that burner will be in use.
* UTILIZE a sparker/lighter with extended nozzle to ignite the Bunsen burner. Never use a match to ignite burner.
* HAVE the sparker/lighter available before turning on gas.
* ADJUST the flame by turning the collar to regulate air flow and produce an appropriate flame for the experiment (typically a medium blue flame).
* DO NOT leave open flames unattended and never leave laboratory while burner is on.
* SHUT-OFF gas when its use is complete.
* ALLOW the burner to cool before handling.
* ENSURE that the main gas valve is off before leaving the laboratory.

Additional Precautions:
* Do not use a Bunsen burner in biological safety cabinet.
* For small fires, attempt to extinguish fire if you been trained in fire extinguisher use.
* In case of a large fire activate the fire alarm, evacuate the building and alert authorities.

==Personal Protective Equipment (PPE)==
Wear standard laboratory attire including safety glasses and avoid wearing synthetic clothing.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

TREATMENT FACILITIES:
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for additional accident and injury reporting information.

==Release/Leak Procedure==
* When a gas leak occurs, personal safety should always come first.
* Alert and clear everyone in the immediate area where the gas leak occurred.
* Open outside windows, if possible & safe to do so.
* Avoid breathing gas.
* Contact OSEH-HMM (734) 763-4568 for release assistance.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when working with Bunsen burners.

Certification
I have read and understand the above SOP. I agree to contact my Supervisor or Lab Manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:
* [[User:Ccousz|Ccousz]] ([[User talk:Ccousz|talk]]) 16:58, 12 October 2016 (UTC)
* [[User:Davebridges|Davebridges]] ([[User talk:Davebridges|talk]]) 14:50, 8 November 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:17, 10 November 2016 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:46, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO

Laboratory Director - Dave Bridges Revision Date - 2016-10-12

SOP - Bleach

2017-06-12T21:45:37Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
This standard operating procedure outlines the handling and use of bleach. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling bleach.
This document is modified from http://www.oseh.umich.edu/docs/SOP/Bleach.docx

Bleach is a general term that typically refers to an aqueous mixture that has a primary active ingredient of either Sodium Hypochlorite or Calcium Hypochlorite and usually at concentrations of 3 - 10%. Bleach is used primarily as an oxidizing, cleaning, or a bleaching agent, as a disinfectant or within drinking water and waste water purification systems.

Synonyms of bleach include sodium hypochlorite, Clorox, liquid bleach, antiformin, chlorox, Carrel-dakin solution, Chloros, Dakin’s solution, hychlorite, Javelle water.

==Useful Bleach Links==
* http://www.atsdr.cdc.gov/toxfaqs/tfacts184.pdf
* http://www.atsdr.cdc.gov/Mhmi/mmg184.pdf
* http://www.asc.co.id/uplimg/File/03%20zMSDS_NaClO_ASC%20R3.pdf
* http://www.cl2.com/documents/Incompatibility%20Chart.pdf

==Potential Hazards==
* Corrosive. May cause severe irritation or damage to eyes or skin. Vapor may irritate eyes and respiratory tract.
* Incompatible with many chemicals/agents. Hazardous gases (including chlorine and chloramines) may be released if bleach is mixed with an incompatible material.
* Exposure to these gases can cause coughing, shortness of breath, irritation to, or burning of the eyes, nose, and throat, chest pain, wheezing, fluid in the lungs, and nausea. Chlorine can also be absorbed through the skin and cause pain, inflammation, swelling, and blistering.
* Strong oxidizer. May initiate or promote combustion in other materials.

==Occupational Exposure Limits (OELs)==
* MIOSHA: 0.5 ppm Chlorine (from sodium hypochlorite), 8-hour PEL
* MIOSHA: 1 ppm Chlorine (from sodium hypochlorite), 15-minute STEL
* AIHA (WEEL): 2 mg/m3 Sodium Hypochlorite, 15-minute STEL

==Engineering Controls==
* An eyewash/drench hose combination unit must be available in the immediate work area for any work with corrosive materials, including bleach.
* If large quantities will be used, a safety shower will also be necessary. Contact OSEH at (734) 647-1143 for a determination of the need for a safety shower if there is not one available.
* Depending on the material’s pH or based on its ability to cause severe tissue damage, e.g., formaldehyde, methylene chloride, methyl ethyl ketone peroxide (MEKP), phenol, etc., the location of the emergency shower and/or emergency eyewash shall be within 25 to 100 feet from the hazardous operation.
* A system of local and/or general exhaust is recommended to keep employee exposures below MIOSHA Permissible Exposure Limits (PEL). Local exhaust ventilation (LEV) is generally preferred because it can control the emissions of the contaminant at its source, preventing dispersion of it into the general work area. Contact OSEH at (734) 647-1143 for a determination of the need for an LEV system, if there is not one available.

==Work Practice Controls==
* ALWAYS review the SDS of both bleach and any chemicals/agents before mixing them with bleach to ensure compatibility. A general list of incompatible materials can be found here.
* NEVER mix bleach with an unknown liquid or unknown residue.
* Do not mix bleach with any compound that is incompatible with oxidizers.
* Purchase bleach in the smallest containers that are practical for lab use.
* Work with the smallest practicable amount and lowest practicable concentration.

==Personal Protective Equipment (PPE)==
In order to select the appropriate PPE for the workplace, a Hazard Assessment is conducted. The hazard assessment determines the hazards and potential hazards associated with a task, machinery, or process. The appropriate PPE for the situation may be subsequently determined. The Hazard Assessment Form may be found here. It may be completed either by the workplace supervisor or the department’s OSEH representative.
* Personal protective equipment is especially important. Wear a buttoned lab coat, safety glasses (that meet the requirements of ANSI/ISEA Z87.1) or goggles if splashing may occur, and gloves for any work with bleach. Depending on the quantities and concentrations used, a face shield, impenetrable, chemical apron and sleeves (or coverall), and special gloves may be recommended.
* Should adequate dilution ventilation or LEV be present, respiratory protection should not be necessary. However, in the absence of these and when MIOSHA PEL, or other published occupational exposures limits (OEL) are anticipated to be exceeded, respiratory protection may be necessary. Contact OSEH at (734) 647-1143 for a determination of the need for a respirator.

==Transportation and Storage==
* Transport concentrated bleach solutions in secondary containment, preferably a polyethylene or other non-reactive acid/solvent bottle carrier.
* Store in well-ventilated areas with secondary containment, such as a non-reactive plastic bin.
* Store below eye level.
* Store away from metal (unless the metal has a corrosion-proof coating), and do not store under the sink.
* Store away from incompatibles and flammable materials. Always review the SDS of other chemicals in the storage area for compatibility with bleach.
* Avoid storing on the floor. If storing on the floor is necessary, use secondary containment.

==Waste Disposal==
If the bleach solution is at a household concentration or less, i.e., a maximum 10% concentration of sodium hypochlorite, and was used for disinfecting, limit discharges down a laboratory sink to less than 1-gallon (approx. 3.8 L). If the concentration was >10% and was used for disinfecting, limit the sink disposal to less than 2-cups (approx. 500 mL) of bleach solution. Also flush the drain with adequate volumes of water.

This does not apply to “waste” bleach, e.g., unused or expired bleach. Any leftover, unused product or expired bleach not used as intended by the manufacturer, or used as a disinfectant, needs be collected by OSEH Hazardous Materials Management (HMM) for proper disposal. Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, and waste collection. Also refer to the OSEH Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==
If the employee is in need of emergency medical attention, call 911 immediately.

In general, flush affected eyes or skin with water for at least 15 minutes, then seek medical attention (see below).
* Eyes: In case of contact, immediately flush eyes with plenty of water for at least 15 minutes. Get medical aid.
* Skin: In case of contact, immediately flush skin with plenty of water. Remove contaminated clothing and shoes. Get medical aid. Wash clothing before reuse.
* Ingestion: Get medical aid immediately. Do not induce vomiting unless directed to do so by medical personnel. Never give anything by mouth to an unconscious person. If vomiting occurs naturally, have victim lean forward. Wash mouth with water, and then give plenty of milk or water to drink and obtain urgent medical attention.
* Inhalation: If inhaled, remove to fresh air. If not breathing, give artificial respiration. Get medical aid.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==Treatment Facilities==
* U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
* University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
* UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

==Spill Procedures==
* When a spill occurs, personal safety should always come first.
* Alert and clear everyone in the immediate area where the spill occurred.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., less than 1 Gallon or 3.5 Liters. A major/large chemical spill requires active assistance from emergency personnel.

For minor (small) spills
* Use proper personal protective equipment as indicated above.
* Absorb the spilled bleach using an absorbent, non-combustible material such as appropriate sorbent pads, sand, or vermiculite. Do not use combustible materials such as sawdust.
* Collect residue, place in container and contact OSEH-HMM (734) 763-4568 for proper disposal.

For major (large) spills
Report large chemical spills greater than 1 gallon or 3.5 liters in corridors or common areas, e.g., hallways, elevators, eating areas, rest rooms, offices, etc., to University of Michigan Division of Public Safety and Security (DPSS) by calling 911.

Additional Spill Response Steps:

===Minor Chemical Spill===
* Alert people in immediate area of spill.
* Open outside windows, if possible.
* Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.
* Avoid breathing vapors from spill.
* Confine spill to as small an area as possible.
* Do not wash spill down the drain.
* Use appropriate spill kits/sorbents to neutralize corrosives and/or absorb spill. Collect contaminated materials and residues and place in container. For powdered chemicals sweep carefully to avoid generation of dust or, if appropriate, use moist sorbent pads or wet the powder with a suitable solvent and then wipe with a dry cloth. Contact OSEH-HMM (734) 763-4568 for proper disposal.
* Clean spill area with water.

===Major Chemical Spill===
* Attend to injured or contaminated persons and remove them from exposure.
* Alert people in the laboratory to evacuate.
* Close doors to affected area.
* Post warnings to keep people from entering the area.
* Have person available that has knowledge of incident and laboratory to assist emergency personnel.

===Additional Spill Links===
* http://www.oseh.umich.edu/pdf/chemspil.pdf
* http://www.oseh.umich.edu/emer-chemical.shtml

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling bleach.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 16:42, 12 October 2016 (UTC)
* [[User:Ccousz|Ccousz]] ([[User talk:Ccousz|talk]])
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:17, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 18:47, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:45, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☒ NO

Laboratory Director - Dave Bridges Revision Date - 2016-07-26

SOP - B-mercaptoethanol

2017-06-12T21:44:05Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety]]
=Description=
This standard operating procedure (SOP) outlines the handling and use of B-mercaptoethanol (BME). Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling BME.

BME is used as an enzyme reactivator in inhibited systems, a reducing agent in the fluorescent reaction of o-phthaldialdehyde, and amino-acids in alkaline media and is also used to dissociate proteins.

=Potential Hazards=
• BME has a very low odor threshold (0.12-0.64 ppm) and smells similar to the odorant used in natural gas. If the odor becomes widespread, people in nearby areas may suspect a natural gas leak. This can lead to calls to the fire department and/or evacuation of the building, which can be inconvenient and disruptive.

• BME can be toxic if ingested and fatal if inhaled or absorbed through the skin.

• Vapors can irritate the eyes, mucous membranes, and respiratory tract. Symptoms of inhalation exposure may include coughing, sore throat, and/or shortness of breath.

• When BME is heated to decomposition, toxic fumes including sulfur oxides and carbon oxides will be emitted.

• BME is combustible as a liquid or vapor.

• Reactions of BME with strong acids or alkali metals will release flammable hydrogen gas.

=Engineering Controls=
ALWAYS work with BME inside a chemical fume hood or 100% exhausted biological safety cabinet (Class II, Type B2).

=Work Practice Controls=
• BME is incompatible with metals, oxidizing agents, acids, alkalis, calcium hypochlorite, aliphatic amines, and isocyanates.

• Purchase and use in the smallest practical quantities for the experiment being performed.

• Know the location of the nearest fire extinguisher before beginning work.

• Eliminate ignition sources such as open flames and hot surfaces.

• Keep containers closed as much as possible when not in use.

• Be aware of skin absorption as a possible route of exposure. Plan work so that minimal glove contact is expected, and purchase appropriate gloves for cleaning up small spills.

• If glove contact occurs, change gloves immediately.

=Personal Protective Equipment (PPE)=
At a minimum, double-glove using nitrile laboratory gloves and wear a lab coat and safety glasses when working with BME. If there is a possibility of splashing, wear chemical splash goggles and/or a face shield.

=Transportation and Storage=
• Do not store near sources of ignition, oxidizing agents, acids, alkaline compounds, or any other incompatible materials.

• Store BME in a sealed secondary container in a well-ventilated area.

• The container must be tightly closed, resealed, and stored upright to avoid leakage.

• Avoid storing on the floor.

• Transport toxic liquids in secondary containment, preferably a polyethylene or other non-reactive acid/solvent bottle carrier.

• Suitable fire control devices (such as fire extinguishers) must be available at locations where flammable or combustible liquids are stored.

=Waste Disposal=
Handle and store hazardous waste following the guidelines above for work practice controls, transportation and storage. Contact OSEH Hazardous Materials Management at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

=Exposures/Unintended Contact=
If the employee is in need of emergency medical attention, call 911 immediately.

==For an actual chemical exposure/injury:==
• In case of skin contact: Flush the skin with copious amounts of water for at least 15 minutes, and then seek medical attention (see below).

• In case of eye contact: Flush contaminated eye(s) immediately with copious amounts of water for at least 15 minutes, and then seek medical attention (see below).

• In case of inhalation: Assist conscious persons to an area with fresh, uncontaminated air, and then seek medical attention (see below).

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to Work Connections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the Work Connections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===

U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm

After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021

University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm

Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320

UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

==Spill Procedure==
• When a spill occurs, personal safety should always come first.

• Alert and clear everyone in the immediate area where the spill occurred.

Employees in the area should be prepared to clean up minor spills confined to the chemical fume hood. Wearing appropriate gloves (e.g., butyl rubber, Silver Shield®, nitrile), splash goggles, lab coat (and impermeable apron, if available), use an inert absorbent material (sorbent pads, vermiculite, dry sand) to clean up the spill. Do not use combustible materials (i.e. saw dust) to absorb spill. Contaminated PPE and clean-up materials must be placed in a sealed container for pick-up by OSEH-HMM (see waste disposal section for more information).

For large spills, or any spills of BME outside of the fume hood, contact the University of Michigan Division of Public Safety and Security (DPSS) by calling 911.
Additional Spill Links:

• http://oseh.umich.edu/hazardous-waste/spill-response/

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

=Training of Personnel=
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling BME.

=Certification=
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure.

*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 19:42, 10 November 2016 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:43, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☐ NO
Laboratory Director Revision Date
�
Major Revisions (Tracking purposes only – Do not print as part of SOP)

SOP - B-mercaptoethanol

2017-06-12T21:43:39Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety]]
=Description=
This standard operating procedure (SOP) outlines the handling and use of B-mercaptoethanol (BME). Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling BME.

BME is used as an enzyme reactivator in inhibited systems, a reducing agent in the fluorescent reaction of o-phthaldialdehyde, and amino-acids in alkaline media and is also used to dissociate proteins.

=Potential Hazards=
• BME has a very low odor threshold (0.12-0.64 ppm) and smells similar to the odorant used in natural gas. If the odor becomes widespread, people in nearby areas may suspect a natural gas leak. This can lead to calls to the fire department and/or evacuation of the building, which can be inconvenient and disruptive.

• BME can be toxic if ingested and fatal if inhaled or absorbed through the skin.

• Vapors can irritate the eyes, mucous membranes, and respiratory tract. Symptoms of inhalation exposure may include coughing, sore throat, and/or shortness of breath.

• When BME is heated to decomposition, toxic fumes including sulfur oxides and carbon oxides will be emitted.

• BME is combustible as a liquid or vapor.

• Reactions of BME with strong acids or alkali metals will release flammable hydrogen gas.

=Engineering Controls=
ALWAYS work with BME inside a chemical fume hood or 100% exhausted biological safety cabinet (Class II, Type B2).

=Work Practice Controls=
• BME is incompatible with metals, oxidizing agents, acids, alkalis, calcium hypochlorite, aliphatic amines, and isocyanates.

• Purchase and use in the smallest practical quantities for the experiment being performed.

• Know the location of the nearest fire extinguisher before beginning work.

• Eliminate ignition sources such as open flames and hot surfaces.

• Keep containers closed as much as possible when not in use.

• Be aware of skin absorption as a possible route of exposure. Plan work so that minimal glove contact is expected, and purchase appropriate gloves for cleaning up small spills.

• If glove contact occurs, change gloves immediately.

=Personal Protective Equipment (PPE)=
At a minimum, double-glove using nitrile laboratory gloves and wear a lab coat and safety glasses when working with BME. If there is a possibility of splashing, wear chemical splash goggles and/or a face shield.

=Transportation and Storage=
• Do not store near sources of ignition, oxidizing agents, acids, alkaline compounds, or any other incompatible materials.

• Store BME in a sealed secondary container in a well-ventilated area.

• The container must be tightly closed, resealed, and stored upright to avoid leakage.

• Avoid storing on the floor.

• Transport toxic liquids in secondary containment, preferably a polyethylene or other non-reactive acid/solvent bottle carrier.

• Suitable fire control devices (such as fire extinguishers) must be available at locations where flammable or combustible liquids are stored.

=Waste Disposal=
Handle and store hazardous waste following the guidelines above for work practice controls, transportation and storage. Contact OSEH Hazardous Materials Management at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

=Exposures/Unintended Contact=
If the employee is in need of emergency medical attention, call 911 immediately.

==For an actual chemical exposure/injury:==
• In case of skin contact: Flush the skin with copious amounts of water for at least 15 minutes, and then seek medical attention (see below).

• In case of eye contact: Flush contaminated eye(s) immediately with copious amounts of water for at least 15 minutes, and then seek medical attention (see below).

• In case of inhalation: Assist conscious persons to an area with fresh, uncontaminated air, and then seek medical attention (see below).

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to Work Connections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the Work Connections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===

U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm

After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021

University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm

Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320

UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

==Spill Procedure==
• When a spill occurs, personal safety should always come first.

• Alert and clear everyone in the immediate area where the spill occurred.

Employees in the area should be prepared to clean up minor spills confined to the chemical fume hood. Wearing appropriate gloves (e.g., butyl rubber, Silver Shield®, nitrile), splash goggles, lab coat (and impermeable apron, if available), use an inert absorbent material (sorbent pads, vermiculite, dry sand) to clean up the spill. Do not use combustible materials (i.e. saw dust) to absorb spill. Contaminated PPE and clean-up materials must be placed in a sealed container for pick-up by OSEH-HMM (see waste disposal section for more information).

For large spills, or any spills of BME outside of the fume hood, contact the University of Michigan Division of Public Safety and Security (DPSS) by calling 911.
Additional Spill Links:

• http://oseh.umich.edu/hazardous-waste/spill-response/

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

=Training of Personnel=
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling BME.

=Certification=
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure.

*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 19:42, 10 November 2016 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☐ NO
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:43, 12 June 2017 (UTC)
Laboratory Director Revision Date
�
Major Revisions (Tracking purposes only – Do not print as part of SOP)

Safety and Animal Training

2017-06-12T21:39:30Z

Elhabbal: /* Lab Standard Operating Procedures */

These are the classes that lab staff need to take at Michigan:

==General Lab Safety==
These are offered through MyLink at https://maislinc.umich.edu/. All lab personnel need to take these

* BLS025w ONLINE REQUIRED General Laboratory Safety Training (online)
* BLS023 REQUIRED Centrifuge Training (classroom)
* BLS013w ONLINE REQUIRED Autoclave Standard Operating Procedures (online)

If you will be working with radioactive materials, you will have to take this course as well:

* RSS006 REQUIRED Radiation Safety Orientation (classroom)

If you will be working with any viruses you will have to take this course as well:

* BLS008 REQUIRED Working Safely with Viral Vectors (classroom)

If you plan to breed animals, you can also take this course:

*ULAM-10125 Breeding Colony Management for Rats and Mice - Recommended (online)

==Animals==
If you will be working with animals you will have to take these classes. They are all offered through ULAM MLearning (https://trainingportal.med.umich.edu/). This requires a levels 2 password.

* ULAM-10000 Orientation (online)
* ULAM-10050 Hazards pt.1 (online)
* ULAM-10055 Hazards pt.2 (classroom)
* ULAM-10100 Intro Mouse/Rat (online)
* ULAM-10131 Animal Room Procedures pt.1 (online)
* ULAM-10132 Animal Room Procedures pt.2 (classroom)
* ULAM-10105 Lab Mouse Workshop (classroom)

==Lab Standard Operating Procedures==
For lab specific safety training please review the SOP's and sign them if you will be using any of them. All lab SOP's are available [[:Category: SOP|here]]

*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:08, 5 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 13:59, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 19:57, 12 June 2017 (UTC)
[[ Category: Lab Safety ]]
[[ Category: Training ]]
[[Category: SOP]]

SOP - Animal Anesthetics

2017-06-12T21:38:57Z

Elhabbal: /* Certification */

[[ Category: SOP ]]
[[ Category: Lab Safety]]

==Description==
This standard operating procedure outlines the handling and use of animal anesthetics including: isoflurane, halothane, enflurane and ether. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls and personal protective equipment when handling animal anesthetics.

==Procedure Location==
The use of animal anesthetics must be performed in an area with good ventilation and controls to capture and exhaust waste anesthetic gases.

==Potential Hazards==
Anesthetic gas and vapor that leaks during medical or research procedures are considered waste anesthetic gases (WAGs). University faculty, staff and students should be aware of the potential risks of WAGs and be advised to take appropriate precautions to reduce exposures. Workers acutely exposed to excessive amounts of anesthetic gas can experience symptoms of drowsiness, headache, nausea, poor judgment and loss of coordination. Chronic symptoms of over-exposure can include liver, kidney and reproductive effects. Safety precautions include the use of an approved gas scavenging system, or using the agent inside a certified chemical fume hood.

The use of ether is not recommended because it is flammable and a mutagen. Be certain that there are no ignition sources present when handling ether. There are restrictions concerning the use of ether with animals. Contact OSEH at (734) 647-1143 concerning the use of ether.

==Engineering Controls==
Anesthetics should not be handled on the bench top without special ventilation or a scavenging system. Anesthetic gas filtering cartridges, snorkel exhaust, fume hoods or other scavenging systems must be used. ULAM provides ventilated procedure rooms designed for use of anesthetic gases in many areas. Fume hoods provide the best protection against exposure to anesthetics in the laboratory and are the preferred ventilation control device when handling greater than 100 cc outside of the original container. Always handle large quantities of ethyl ether in a fume hood due to the flammable nature of the material. If your research does not permit the handling of large quantities of ethyl ether in your fume hood, contact OSEH to review the adequacy of all special ventilation.

Liquid anesthetics administered with a vaporizer must be scavenged. When used properly, vaporizers equipped with activated charcoal canisters (e.g. F/Air) are effective in removing halogenated waste gases. The F/Air canister containing activated charcoal will absorb waste anesthetics for about 12 hours. Note: F/Air Canisters only absorb halogenated anesthetics (e.g. isoflurane, halothane). Immediately before using any anesthesia machine, the F/Air canister should be removed and weighed to evaluate the remaining absorption capacity. The weight should be recorded and dated on the side of the canister. Immediately following the use of an anesthesia machine, the number of hours the machine was in use should be recorded next to the dated weight information.

Canisters that exceed 12 hours of use or 50 grams of accumulated weight (whichever comes first) must be removed and placed in a sealed plastic bag and disposed of as a hazardous waste through OSEH Hazardous Materials Management (HMM) at (734) 763-4568. Thoroughly clean the induction chamber immediately after each use to avoid residual anesthetic waste release into the environment (which can continue to be released for up to three hours). Please refer to the OSEH Guideline entitled Anesthetic Gas Use for additional information on the safe use of anesthetic gases.

==Work Practice Controls==
All anesthetic agents must be clearly labeled with the correct chemical name. Handwritten labels are acceptable; chemical formulas and structural formulas are not acceptable.

Always keep the flow rate of anesthetics to the animal as low as possible during the procedure. High flow rates can increase your exposure to the anesthetic. It is also important to move the point of potential gas release as close to the exhaust system as possible to increase capture of the chemical.

Do not permit containers to remain open on the bench top. The odor thresholds for most liquid anesthetics (except for ether) are well above permissible exposure limits. If you smell the anesthetic the control procedures you are using are inadequate and must be re-evaluated.

==Personal Protective Equipment (PPE)==
Eye protection in the form of safety glasses must be worn at all times when handling anesthetic agents. Ordinary (street) prescription glasses do not provide adequate protection

Single use nitrile or latex gloves must be worn when handling anesthetic agents as well as lab coats, closed toed shoes and pants. Additional protective clothing should be worn if the possibility of skin contact is likely.

==Transportation and Storage==
Ethers form potentially explosive peroxides after exposure to air and light. Since these chemicals are packaged in an air atmosphere, peroxides can form even though the containers have not been opened. Write the date received and date opened on all containers of ether. Opened containers of ether should be discarded within 12 months of opening. Even closed containers of ether must be discarded by the expiration date through OSEH-HMM (734) 763-4568.

Halogenated liquid anesthetic agents (i.e. halothane, enflurane, isoflurane) are not flammable but do have limited shelf life. Be certain to date the chemical when it is opened and to check expiration date before use.

Always purchase the smallest quantity required for use. Ether used for anesthetic purposes should be purchased in the smallest quantity available (typically 150 cc, Fisher Scientific E136-150) due to its short (12 month) shelf life after it is opened.

==Waste Disposal==
Anesthetic agents are hazardous wastes. Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

'''If the employee is in need of emergency medical attention, call 911 immediately.'''

Wash hands and arms with soap and water immediately following any skin contact with anesthetic agents. Flush eyes for 15 minutes following eye contact.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==Treatment Facilities:==
*U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
*University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
*UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

Click here for additional accident and injury reporting information.

==Spill Procedure==
Ether is extremely flammable. If ether is spilled immediately assess and deactivate potential ignition sources. Be prepared for a potential fire and ensure your safety and others first.

Anticipate spills by having the appropriate clean up equipment on hand. Spill materials for anesthetic agents are designed to control the liquid portion of the spill and minimize the production of vapors. Never use paper towels on large spills of anesthetic agents because it exacerbates vapor production.

*When a spill occurs, personal safety should always come first.
*Alert and clear everyone in the immediate area where the spill occurred.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., less than 1 liter. A major/large chemical spill requires active assistance from emergency personnel.

==Additional Spill Response Steps:==

===MINOR CHEMICAL SPILL===
*Alert people in immediate area of spill.
*If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
*Open outside windows, if possible.
*Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.
*Avoid breathing vapors from spill.
*Confine spill to as small an area as possible.
*Do not wash spill down the drain.
*Use appropriate spill kits/sorbents to absorb spill. Collect contaminated materials and residues and place in container. Contact OSEH-HMM (734) 763-4568 for proper disposal.
*Clean spill area with water.

===MAJOR CHEMICAL SPILL===
*Attend to injured or contaminated persons and remove them from exposure.
*Alert people in the laboratory to evacuate.
*If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
*Call University of Michigan Division of Public Safety and Security (DPSS) at 911 immediately for assistance.
*Close doors to affected area.
*Post warnings to keep people from entering the area.
*Have person available that has knowledge of incident and laboratory to assist emergency personnel.

==Additional Spill Links:==
• www.oseh.umich.edu/pdf/chemspil.pdf
• http://www.oseh.umich.edu/emer-chemical.shtml.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling animal anesthetics.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:44, 13 October 2016 (UTC)

* [[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 19:34, 10 November 2016 (UTC)

*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:09, 5 June 2017 (UTC)

* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 18:34, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:38, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO

Laboratory Director - Dave Bridges Revision Date - 2016-10-13

SOP - Animal Anesthetics

2017-06-12T21:38:24Z

Elhabbal:

[[ Category: SOP ]]
[[ Category: Lab Safety]]

==Description==
This standard operating procedure outlines the handling and use of animal anesthetics including: isoflurane, halothane, enflurane and ether. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls and personal protective equipment when handling animal anesthetics.

==Procedure Location==
The use of animal anesthetics must be performed in an area with good ventilation and controls to capture and exhaust waste anesthetic gases.

==Potential Hazards==
Anesthetic gas and vapor that leaks during medical or research procedures are considered waste anesthetic gases (WAGs). University faculty, staff and students should be aware of the potential risks of WAGs and be advised to take appropriate precautions to reduce exposures. Workers acutely exposed to excessive amounts of anesthetic gas can experience symptoms of drowsiness, headache, nausea, poor judgment and loss of coordination. Chronic symptoms of over-exposure can include liver, kidney and reproductive effects. Safety precautions include the use of an approved gas scavenging system, or using the agent inside a certified chemical fume hood.

The use of ether is not recommended because it is flammable and a mutagen. Be certain that there are no ignition sources present when handling ether. There are restrictions concerning the use of ether with animals. Contact OSEH at (734) 647-1143 concerning the use of ether.

==Engineering Controls==
Anesthetics should not be handled on the bench top without special ventilation or a scavenging system. Anesthetic gas filtering cartridges, snorkel exhaust, fume hoods or other scavenging systems must be used. ULAM provides ventilated procedure rooms designed for use of anesthetic gases in many areas. Fume hoods provide the best protection against exposure to anesthetics in the laboratory and are the preferred ventilation control device when handling greater than 100 cc outside of the original container. Always handle large quantities of ethyl ether in a fume hood due to the flammable nature of the material. If your research does not permit the handling of large quantities of ethyl ether in your fume hood, contact OSEH to review the adequacy of all special ventilation.

Liquid anesthetics administered with a vaporizer must be scavenged. When used properly, vaporizers equipped with activated charcoal canisters (e.g. F/Air) are effective in removing halogenated waste gases. The F/Air canister containing activated charcoal will absorb waste anesthetics for about 12 hours. Note: F/Air Canisters only absorb halogenated anesthetics (e.g. isoflurane, halothane). Immediately before using any anesthesia machine, the F/Air canister should be removed and weighed to evaluate the remaining absorption capacity. The weight should be recorded and dated on the side of the canister. Immediately following the use of an anesthesia machine, the number of hours the machine was in use should be recorded next to the dated weight information.

Canisters that exceed 12 hours of use or 50 grams of accumulated weight (whichever comes first) must be removed and placed in a sealed plastic bag and disposed of as a hazardous waste through OSEH Hazardous Materials Management (HMM) at (734) 763-4568. Thoroughly clean the induction chamber immediately after each use to avoid residual anesthetic waste release into the environment (which can continue to be released for up to three hours). Please refer to the OSEH Guideline entitled Anesthetic Gas Use for additional information on the safe use of anesthetic gases.

==Work Practice Controls==
All anesthetic agents must be clearly labeled with the correct chemical name. Handwritten labels are acceptable; chemical formulas and structural formulas are not acceptable.

Always keep the flow rate of anesthetics to the animal as low as possible during the procedure. High flow rates can increase your exposure to the anesthetic. It is also important to move the point of potential gas release as close to the exhaust system as possible to increase capture of the chemical.

Do not permit containers to remain open on the bench top. The odor thresholds for most liquid anesthetics (except for ether) are well above permissible exposure limits. If you smell the anesthetic the control procedures you are using are inadequate and must be re-evaluated.

==Personal Protective Equipment (PPE)==
Eye protection in the form of safety glasses must be worn at all times when handling anesthetic agents. Ordinary (street) prescription glasses do not provide adequate protection

Single use nitrile or latex gloves must be worn when handling anesthetic agents as well as lab coats, closed toed shoes and pants. Additional protective clothing should be worn if the possibility of skin contact is likely.

==Transportation and Storage==
Ethers form potentially explosive peroxides after exposure to air and light. Since these chemicals are packaged in an air atmosphere, peroxides can form even though the containers have not been opened. Write the date received and date opened on all containers of ether. Opened containers of ether should be discarded within 12 months of opening. Even closed containers of ether must be discarded by the expiration date through OSEH-HMM (734) 763-4568.

Halogenated liquid anesthetic agents (i.e. halothane, enflurane, isoflurane) are not flammable but do have limited shelf life. Be certain to date the chemical when it is opened and to check expiration date before use.

Always purchase the smallest quantity required for use. Ether used for anesthetic purposes should be purchased in the smallest quantity available (typically 150 cc, Fisher Scientific E136-150) due to its short (12 month) shelf life after it is opened.

==Waste Disposal==
Anesthetic agents are hazardous wastes. Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

'''If the employee is in need of emergency medical attention, call 911 immediately.'''

Wash hands and arms with soap and water immediately following any skin contact with anesthetic agents. Flush eyes for 15 minutes following eye contact.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==Treatment Facilities:==
*U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
*University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
*UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

Click here for additional accident and injury reporting information.

==Spill Procedure==
Ether is extremely flammable. If ether is spilled immediately assess and deactivate potential ignition sources. Be prepared for a potential fire and ensure your safety and others first.

Anticipate spills by having the appropriate clean up equipment on hand. Spill materials for anesthetic agents are designed to control the liquid portion of the spill and minimize the production of vapors. Never use paper towels on large spills of anesthetic agents because it exacerbates vapor production.

*When a spill occurs, personal safety should always come first.
*Alert and clear everyone in the immediate area where the spill occurred.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., less than 1 liter. A major/large chemical spill requires active assistance from emergency personnel.

==Additional Spill Response Steps:==

===MINOR CHEMICAL SPILL===
*Alert people in immediate area of spill.
*If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
*Open outside windows, if possible.
*Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.
*Avoid breathing vapors from spill.
*Confine spill to as small an area as possible.
*Do not wash spill down the drain.
*Use appropriate spill kits/sorbents to absorb spill. Collect contaminated materials and residues and place in container. Contact OSEH-HMM (734) 763-4568 for proper disposal.
*Clean spill area with water.

===MAJOR CHEMICAL SPILL===
*Attend to injured or contaminated persons and remove them from exposure.
*Alert people in the laboratory to evacuate.
*If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
*Call University of Michigan Division of Public Safety and Security (DPSS) at 911 immediately for assistance.
*Close doors to affected area.
*Post warnings to keep people from entering the area.
*Have person available that has knowledge of incident and laboratory to assist emergency personnel.

==Additional Spill Links:==
• www.oseh.umich.edu/pdf/chemspil.pdf
• http://www.oseh.umich.edu/emer-chemical.shtml.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling animal anesthetics.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:44, 13 October 2016 (UTC)

* [[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 19:34, 10 November 2016 (UTC)

*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:09, 5 June 2017 (UTC)

* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 18:34, 7 June 2017 (UTC)
[[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:38, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO

Laboratory Director - Dave Bridges Revision Date - 2016-10-13

Safety and Animal Training

2017-06-12T19:57:24Z

Elhabbal:

These are the classes that lab staff need to take at Michigan:

==General Lab Safety==
These are offered through MyLink at https://maislinc.umich.edu/. All lab personnel need to take these

* BLS025w ONLINE REQUIRED General Laboratory Safety Training (online)
* BLS023 REQUIRED Centrifuge Training (classroom)
* BLS013w ONLINE REQUIRED Autoclave Standard Operating Procedures (online)

If you will be working with radioactive materials, you will have to take this course as well:

* RSS006 REQUIRED Radiation Safety Orientation (classroom)

If you will be working with any viruses you will have to take this course as well:

* BLS008 REQUIRED Working Safely with Viral Vectors (classroom)

If you plan to breed animals, you can also take this course:

*ULAM-10125 Breeding Colony Management for Rats and Mice - Recommended (online)

==Animals==
If you will be working with animals you will have to take these classes. They are all offered through ULAM MLearning (https://trainingportal.med.umich.edu/). This requires a levels 2 password.

* ULAM-10000 Orientation (online)
* ULAM-10050 Hazards pt.1 (online)
* ULAM-10055 Hazards pt.2 (classroom)
* ULAM-10100 Intro Mouse/Rat (online)
* ULAM-10131 Animal Room Procedures pt.1 (online)
* ULAM-10132 Animal Room Procedures pt.2 (classroom)
* ULAM-10105 Lab Mouse Workshop (classroom)

==Lab Standard Operating Procedures==
For lab specific safety training please review the SOP's and sign them if you will be using any of them. All lab SOP's are available [[:Category: SOP|here]]

*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:08, 5 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 13:59, 9 June 2017 (UTC)
[[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 19:57, 12 June 2017 (UTC)
[[ Category: Lab Safety ]]
[[ Category: Training ]]
[[Category: SOP]]

Splitting Cells

2017-06-01T17:31:25Z

Elhabbal:

==Materials==
*Media (FBS or NCS as required): Prepare by filtering together 5 mL PSG, 50 mL COS-FBS and one bottle of DMEM
*PBS -/-
*0.05% Trypsin

==Protocol==
#Warm PBS and Media in water bath
# Aspirate the plate media
#Wash cells once with 10 mL (per 10 cm dish) PBS -/- then aspirate the PBS
#Add 1 mL trypsin and allow to sit in the hood for 2-5 min
#Add 10 mL media to each new dish
#Check cells for trypsinization, and if necessary tap the cells
#Add 9 mL media to trypsinized cells
#Add 1 mL cells to each dish (for 10X dilution; 0.5 mL for 20X)
#Replace plates in the 37C incubator

==Cell Specific Notes==
*3T3-L1 fibroblasts have special considerations regarding confluence. See [[Differentiation of 3T3-L1 Cells]]
*RAW 264.7 cells are scraped, not trypsinized. See [[Culturing RAW 264.7 Cells]]
*S2 cells are grown at 28C without extra CO2. See [[Culturing S2 Cells]]
[[ Category:Cell Culture ]]

Splitting Cells

2017-06-01T17:28:28Z

Elhabbal: /* Protocol */ added the aspiration processes and changed wash cells twice to "once"

==Materials==
*Media (FBS or NCS as required): Prepare by filtering together 5 mL PSG, 50 mL COS-FBS and one bottle of DMEM
*PBS -/-
*0.05% Trypsin

==Protocol==
#Warm PBS and Media in water bath
# Aspirate media
#Wash cells once with 10 mL (per 10 cm dish) PBS -/- then aspirate the PBS
#Add 1 mL trypsin and allow to sit in the hood for 2-5 min
#Add 10 mL media to each new dish
#Check cells for trypsinization, and if necessary tap the cells
#Add 9 mL media to trypsinized cells
#Add 1 mL cells to each dish (for 10X dilution; 0.5 mL for 20X)
#Replace plates in the 37C incubator

==Cell Specific Notes==
*3T3-L1 fibroblasts have special considerations regarding confluence. See [[Differentiation of 3T3-L1 Cells]]
*RAW 264.7 cells are scraped, not trypsinized. See [[Culturing RAW 264.7 Cells]]
*S2 cells are grown at 28C without extra CO2. See [[Culturing S2 Cells]]
[[ Category:Cell Culture ]]

Safety and Animal Training

2017-05-30T03:23:30Z

Elhabbal: added "(classroom)" to BLS 008 course

These are the classes that lab staff need to take at Michigan:

==General Lab Safety==
These are offered through MyLink at https://maislinc.umich.edu/. All lab personnel need to take these

* BLS025w ONLINE REQUIRED General Laboratory Safety Training (online)
* BLS023 REQUIRED Centrifuge Training (classroom)
* BLS013w ONLINE REQUIRED Autoclave Standard Operating Procedures (online)

If you will be working with radioactive materials, you will have to take this course as well:

* RSS006 REQUIRED Radiation Safety Orientation (classroom)

If you will be working with any viruses you will have to take this course as well:

* BLS008 REQUIRED Working Safely with Viral Vectors (classroom)

If you plan to breed animals, you can also take this course:

*ULAM-10125 Breeding Colony Management for Rats and Mice - Recommended (online)

==Animals==
If you will be working with animals you will have to take these classes. They are all offered through ULAM MLearning (https://trainingportal.med.umich.edu/). This requires a levels 2 password.

* ULAM-10000 Orientation (online)
* ULAM-10050 Hazards pt.1 (online)
* ULAM-10055 Hazards pt.2 (classroom)
* ULAM-10100 Intro Mouse/Rat (online)
* ULAM-10131 Animal Room Procedures pt.1 (online)
* ULAM-10132 Animal Room Procedures pt.2 (classroom)
* ULAM-10105 Lab Mouse Workshop (classroom)

==Lab Standard Operating Procedures==
For lab specific safety training please review the SOP's and sign them if you will be using any of them. All lab SOP's are available [[:Category: SOP|here]]

*[[User:Lpfeifer|Lpfeifer]] ([[User talk:Lpfeifer|talk]]) 14:43, 10 November 2016 (UTC)

[[ Category: Lab Safety ]]
[[ Category: Training ]]
[[Category: SOP]]