Bridges Lab Protocols - User contributions [en]

Osmotic Minipump Implantation

2019-12-04T21:55:51Z

Snyderds:

This page is a repository of resources for using the ALZET osmotic minipump.

Before selecting a pump or planning an experiment all users should first explore the pages under the "http://www.alzet.com products>Guide to Use"] dropdown menus with particular attention to the following pages to assure appropriate selection and application of a pump for an experimental study:

*[https://www.alzet.com/guide-to-use/checklist-and-tips-for-successful-use-of-alzet-osmotic-pumps/ Manufacturer’s comprehensive checklist]
*[https://www.alzet.com/resources/alzet-technical-tips/ Technical Tips]
*[https://www.alzet.com/alzaid-chemical-compatibility-test-kit/ Chemical compatibility]
*[https://www.alzet.com/guide-to-use/implementation_and_explanation/ Minipump implantation Suggestions]
*[https://www.alzet.com/guide-to-use/implementation_and_explanation/ Surgical instruction manual]
*[https://vimeopro.com/user38667762/alzet-resource-videos ALZET resources Vimeo channel]
*https://www.alzet.com/products/alzet_pumps/specifications/ ALZET Product Rate and Durations Specifications]
*[https://www.alzet.com/guide-to-use/filling-priming-alzet-pumps/ Pump Filling and Priming Instructions]
*[https://www.alzet.com/guide-to-use/mri/ Use of osmotic pumps with MRI]
*[https://www.alzet.com/formulating-the-solution/ Concentration calculator]

For details on study design considerations (protocols, timeline, power calculations, etc.) as developed in pilot leptin infusion experiments, [https://docs.google.com/presentation/d/15R6rhEefvqhxJfwDy019DONKbSbp6x27wdQf4gyGYsA/edit?usp=sharing pleaser refer to this shared slide deck].

For additional resources, you may refer to [https://drive.google.com/open?id=1wubhf3yG4VY4SiBqLbfKmm8myrwXt_Ts this shared google drive folder].

Consider setting up a consultation with ULAM veterinary services for a demonstration and practice before performing your own surgery.

'''Pre-surgery checklist:
'''

☐ Stack of paper towels, gauze, q-tips

☐ Heating pad

☐ Sterile forceps (x2)

☐ Hemostat

☐ Incisors

☐ Wound clips

☐ Wound clip applicator and wound clip remover

☐ Electric razor (alternately: Nair)

☐ Lint roller

☐ Isoflurane pump

☐ Eye lubricant

☐ Surgery wash and rinse

☐ ALZET osmotic pumps, filled and primed ahead of time

☐ Post-operative analgesic (ie carprofen)

'''Helpful Tips'''
*'''The ALZET customer support team is EXTREMELY helpful if you are experiencing issues.''' [mailto:alzet@durect.com alzet@durect.com] or 800-692-2990
*Use of a sterile filter may not be necessary when using extremely pure solutes so long as you utilize otherwise aseptic technique. This minimizes waste of costly solutes.
*When using the smaller pump sizes, special considerations for MRI may be disregarded.
*When filling the pumps, the syringe needle needs to be a fraction of a millimeter smaller than the hole in the pump in which it is inserted. This means that you will likely have better success using an '''insulin syringe''', which, owing to its small gauge, allows displaced air to escape when filling.
*Here are some ULAM Surgery guidelines [https://azanimal.sites.uofmhosting.net/node/58 Guidelines on Administration of Substances to Laboratory Animals] and [https://az.research.umich.edu/animalcare/guidelines/guidelines-performance-surgery-rodents ULAM Anesthesia and Analgesia Guidelines]
*Document the rodent surgery recovery with [https://azanimal.sites.uofmhosting.net/node/36 Rodent Surgery Record] and [https://azanimal.sites.uofmhosting.net/node/36 Cage Card Form Rodent Surgical Record].

Osmotic Minipump Implantation

2019-12-04T21:53:50Z

Snyderds:

This page is a repository of resources for using the ALZET osmotic minipump.

Before selecting a pump or planning an experiment all users should first explore the pages under the "http://www.alzet.com products>Guide to Use"] dropdown menus with particular attention to the following pages to assure appropriate selection and application of a pump for an experimental study:

*[https://www.alzet.com/guide-to-use/checklist-and-tips-for-successful-use-of-alzet-osmotic-pumps/ Manufacturer’s comprehensive checklist]
*[https://www.alzet.com/resources/alzet-technical-tips/ Technical Tips]
*[https://www.alzet.com/alzaid-chemical-compatibility-test-kit/ Chemical compatibility]
*[https://www.alzet.com/guide-to-use/implementation_and_explanation/ Minipump implantation Suggestions]
*[https://www.alzet.com/guide-to-use/implementation_and_explanation/ Surgical instruction manual]
*[https://vimeopro.com/user38667762/alzet-resource-videos ALZET resources Vimeo channel]
*https://www.alzet.com/products/alzet_pumps/specifications/ ALZET Product Rate and Durations Specifications]
*[https://www.alzet.com/guide-to-use/filling-priming-alzet-pumps/ Pump Filling and Priming Instructions]
*[https://www.alzet.com/guide-to-use/mri/ Use of osmotic pumps with MRI]
*[https://www.alzet.com/formulating-the-solution/ Concentration calculator]

For details on study design considerations (protocols, timeline, power calculations, etc.) as developed in pilot leptin infusion experiments, [https://docs.google.com/presentation/d/15R6rhEefvqhxJfwDy019DONKbSbp6x27wdQf4gyGYsA/edit?usp=sharing pleaser refer to this shared slide deck].

For additional resources, you may refer to [https://drive.google.com/open?id=1wubhf3yG4VY4SiBqLbfKmm8myrwXt_Ts this shared google drive folder].

Consider setting up a consultation with ULAM veterinary services for a demonstration and practice before performing your own surgery.

'''Pre-surgery checklist:
'''

☐ Stack of paper towels, gauze, q-tips

☐ Heating pad

☐ Sterile forceps (x2)

☐ Hemostat

☐ Incisors

☐ Wound clips

☐ Wound clip applicator and wound clip remover

☐ Electric razor (alternately: Nair)

☐ Lint roller

☐ Isoflurane pump

☐ Eye lubricant

☐ Surgery wash and rinse

☐ ALZET osmotic pumps, filled and primed ahead of time

☐ Post-operative analgesic (ie carprofen)

'''Helpful Tips'''
*'''The ALZET customer support team is EXTREMELY helpful if you are experiencing issues.''' [mailto:alzet@durect.com alzet@durect.com] or 800-692-2990
*Use of a sterile filter may not be necessary when using extremely pure solutes so long as you utilize otherwise aseptic technique. This minimizes waste of costly solutes.
*When using the smaller pump sizes, special considerations for MRI may be disregarded.
*When filling the pumps, the syringe needle needs to be a fraction of a millimeter smaller than the hole in the pump in which it is inserted. This means that you will likely have better success using an '''insulin syringe''', which, owing to its small gauge, allows displaced air to escape when filling.
*Document the rodent surgery recovery with [[https://azanimal.sites.uofmhosting.net/node/36 Rodent Surgery Record]] and [[https://azanimal.sites.uofmhosting.net/node/36 Cage Card Form Rodent Surgical Record]].

Osmotic Minipump Implantation

2019-11-18T23:44:58Z

Snyderds: created page

Ketone Body Tolerance Test

2019-08-08T16:49:37Z

Snyderds:

'''Materials'''

*Ketometer - Precision xtra ketone body / glucose monitor (Abbot)
*Ketone Body Test Strips
*Scale
*Bowl for weighing mice
*echoMRI if the mice differ in body fat levels (see below)
*Syringes
*1.5 mg/mL bOHB in PBS (make as 15g bOHB in 10mL of PBS, sterile filtered). This will correspond to 1 U/kg injections. If you are using a higher or lower dose of bOHB, add more or less to the 10 mL of PBS, so that injections are 10 uL/g of mass.
*This may need to be adjusted depending on the ketone sensitivity of the mice, and this is based on PGC-1a on chow.
*In general you want the bOHB to increase blood bOHB levels by about 10 to 12-fold (bringing bOHB levels form around .5mmol/L up to the 5 to 6 mmol/L range if fasted or 1.5 to 2.5 mmol/L if fed in the most responsive of your two groups. If your response is lower or higher, you will probably have to change your dose and retry.
*Timer

'''Protocol'''

*'''If''' testing fasted ketone tolerance: Remove food from mice for about 6h by putting them in a fresh cage. Add “do not feed” acetate to cages, and ideally move cage to procedure room. Try to make sure that the mice are in a quiet, undisturbed temperature controlled room with the lights on. Typically starve the mice at 8AM and aim to start injections at 2PM
*Prepare a 1 g/10mL solution of glucose in the unlikely case that some animals become hyperketonemic.
*Weigh/MRI mice ahead of time, mark tails if necessary with different colors for rapid identification and take fasting ketone measurement via a tail clip.
*Prepare bOHB syringes with 1.5 g/KG mouse weight (ie for a mouse with 15g lean body mass, use 15uL).
*Pre-open enough ketone test strips for one round of measurements.
*At approximately 40 sec to 1 min intervals, inject appropriate amount of bOHB into intraperitoneal cavity of the mouse.
**Immobilize mouse and restrain tail with one hand.
**Aim needle for peritoneal space, between the midline and the hip bone.
**Insert syringe (do not inject) into cavity.
**Eject syringe.
*At 15 minute intervals (normally 15, 30, 45, 60, 75, & 90 min), take blood bOHB measurements from tail vein. If needed re-snip the tail tip to expose the vein. When measuring BOHB just lift the tail of the mouse, while leaving it in the cage, rather than removing and restraining the mouse which can be more stressful.
*Analyze data by both % change from fasting ketones and absolute values. Our preferred outcome is to report fasting ketone levels and report percent change as a figure.
*The preferred statistical model is a mixed linear model using the time points as ordinate values and testing for a main effect or an interaction of the treatment/genotype. Use the R lme4 package for this.

[[Category:Ketogenesis]]

Ketone Body Tolerance Test

2019-07-30T16:32:23Z

Snyderds:

'''Materials'''

*Ketometer - Precision xtra ketone body / glucose monitor (Abbot)
*Ketone Body Test Strips
*Scale
*Bowl for weighing mice
*echoMRI if the mice differ in body fat levels (see below)
*Syringes
*1.5 mg/mL bOHB in PBS (make as 15g bOHB in 10mL of PBS, sterile filtered). This will correspond to 1 U/kg injections. If you are using a higher or lower dose of bOHB, add more or less to the 10 mL of PBS, so that injections are 10 uL/g of mass.
*This may need to be adjusted depending on the ketone sensitivity of the mice, and this is based on PGC-1a on chow.
*In general you want the bOHB to increase blood bOHB levels by about 10 to 12-fold (bringing bOHB levels form around .5mmol/L up to the 5 to 6 mmol/L range in the most responsive of your two groups. If your response is lower or higher, you will probably have to change your dose and retry.
*Timer

'''Protocol'''

*If testing fasted ketone tolerance: Remove food from mice for about 6h by putting them in a fresh cage. Add “do not feed” acetate to cages, and ideally move cage to procedure room. Try to make sure that the mice are in a quiet, undisturbed temperature controlled room with the lights on.
**Typically starve the mice at 8AM and aim to start injections at 2PM
*Prepare a 1 g/10mL solution of glucose in the unlikely case that some animals become hyperketonemic.
*Weigh/MRI mice ahead of time, mark tails if necessary with different colors for rapid identification and take fasting ketone measurement via a tail clip.
*Prepare bOHB syringes with 1.5 g/KG mouse weight (ie for a mouse with 15g lean body mass, use 15uL).
*At approximately 1 min intervals, inject appropriate amount of bOHB into intraperitoneal cavity of the mouse.
**Immobilize mouse and restrain tail with one hand.
**Aim needle for peritoneal space, between the midline and the hip bone.
**Insert syringe (do not inject) into cavity.
**Eject syringe.
*At 15 minute intervals (normally 15, 30, 45, 60, 75, & 90 min), take blood bOHB measurements from tail vein. If needed re-snip the tail tip to expose the vein. When measuring BOHB just lift the tail of the mouse, while leaving it in the cage, rather than removing and restraining the mouse which can be more stressful.
*Analyze data by both % change from fasting ketones and absolute values. Our preferred outcome is to report fasting ketone levels and report percent change as a figure.
*The preferred statistical model is a mixed linear model using the time points as ordinate values and testing for a main effect or an interaction of the treatment/genotype. Use the R lme4 package for this.

[[Category:Ketogenesis]]

Ketone Body Tolerance Test

2019-07-26T13:40:02Z

Snyderds:

'''Materials'''

*Ketometer - Precision xtra ketone body / glucose monitor (Abbot)
*Ketone Body Test Strips
*Scale
*Bowl for weighing mice
*echoMRI if the mice differ in body fat levels (see below)
*Syringes
*1.5 mg/mL bOHB in PBS (make as 15mg bOHB in 10mL of PBS, sterile filtered). This will correspond to 1 U/kg injections. If you are using a higher or lower dose of bOHB, add more or less to the 10 mL of PBS, so that injections are 10 uL/g of mass.
*This may need to be adjusted depending on the ketone sensitivity of the mice, and this is based on PGC-1a on chow.
*In general you want the bOHB to increase blood bOHB levels by about 10 to 12-fold (bringing bOHB levels form around .5mmol/L up to the 5 to 6 mmol/L range in the most responsive of your two groups. If your response is lower or higher, you will probably have to change your dose and retry.
*Timer

'''Protocol'''

*If testing fasted ketone tolerance: Remove food from mice for about 6h by putting them in a fresh cage. Add “do not feed” acetate to cages, and ideally move cage to procedure room. Try to make sure that the mice are in a quiet, undisturbed temperature controlled room with the lights on.
**Typically starve the mice at 8AM and aim to start injections at 2PM
*Prepare a 1 g/10mL solution of glucose in the unlikely case that some animals become hyperketonemic.
*Weigh/MRI mice ahead of time, mark tails if necessary with different colors for rapid identification and take fasting ketone measurement via a tail clip.
*Prepare bOHB syringes with 1.5 g/KG mouse weight (ie for a mouse with 15g lean body mass, use 15uL).
*At approximately 1 min intervals, inject appropriate amount of bOHB into intraperitoneal cavity of the mouse.
**Immobilize mouse and restrain tail with one hand.
**Aim needle for peritoneal space, between the midline and the hip bone.
**Insert syringe (do not inject) into cavity.
**Eject syringe.
*At 15 minute intervals (normally 15, 30, 45, 60, 75, & 90 min), take blood bOHB measurements from tail vein. If needed re-snip the tail tip to expose the vein. When measuring BOHB just lift the tail of the mouse, while leaving it in the cage, rather than removing and restraining the mouse which can be more stressful.
*Analyze data by both % change from fasting ketones and absolute values. Our preferred outcome is to report fasting ketone levels and report percent change as a figure.
*The preferred statistical model is a mixed linear model using the time points as ordinate values and testing for a main effect or an interaction of the treatment/genotype. Use the R lme4 package for this.

[[Category:Ketogenesis]]

Ketone Body Tolerance Test

2019-07-26T13:21:10Z

Snyderds:

'''Materials'''

*Ketometer - Precision xtra ketone body / glucose monitor (Abbot)
*Ketone Body Test Strips
*Scale
*Bowl for weighing mice
*echoMRI if the mice differ in body fat levels (see below)
*Syringes
*1.5 mg/mL bOHB in PBS (make as 15mg bOHB in 10mL of PBS, sterile filtered). This will correspond to 1 U/kg injections. If you are using a higher or lower dose of bOHB, add more or less to the 10 mL of PBS, so that injections are 10 uL/g of mass.
*This may need to be adjusted depending on the ketone sensitivity of the mice, and this is based on PGC-1a on chow.
*In general you want the bOHB to increase blood bOHB levels by about 10 to 12-fold (bringing bOHB levels form around .5mmol/L up to the 5 to 6 mmol/L range in the most responsive of your two groups. If your response is lower or higher, you will probably have to change your dose and retry.
*Timer

'''Protocol'''

*If testing fasted ketone tolerance: Remove food from mice for about 6h by putting them in a fresh cage. Add “do not feed” acetate to cages, and ideally move cage to procedure room. Try to make sure that the mice are in a quiet, undisturbed temperature controlled room with the lights on.
**Typically starve the mice at 8AM and aim to start injections at 2PM
*Prepare a 1 g/10mL solution of glucose in the unlikely case that some animals become hyperketonemic.
*Weigh/MRI mice ahead of time, mark tails if necessary with different colors for rapid identification and take fasting ketone measurement via a tail clip.
*Prepare bOHB syringes with 1.5 g/KG mouse weight (ie for a 30g mouse, 300 uL).
*At approximately 1 min intervals, inject appropriate amount of bOHB into intraperitoneal cavity of the mouse.
**Immobilize mouse and restrain tail with one hand.
**Aim needle for peritoneal space, between the midline and the hip bone.
**Insert syringe (do not inject) into cavity.
**Eject syringe.
*At 15 minute intervals (normally 15, 30, 45, 60, 75, & 90 min), take blood bOHB measurements from tail vein. If needed re-snip the tail tip to expose the vein. When measuring BOHB just lift the tail of the mouse, while leaving it in the cage, rather than removing and restraining the mouse which can be more stressful.
*Analyze data by both % change from fasting ketones and absolute values. Our preferred outcome is to report fasting ketone levels and report percent change as a figure.
*The preferred statistical model is a mixed linear model using the time points as ordinate values and testing for a main effect or an interaction of the treatment/genotype. Use the R lme4 package for this.
[[Category:Ketogenesis]]

Ketone Body Tolerance Test

2019-07-26T13:16:20Z

Snyderds: Created page with "'''Materials''' *Ketometer - Precision xtra ketone body / glucose monitor (Abbot) *Ketone Body Test Strips *Scale *Bowl for weighing mice *echoMRI if the mice differ in body..."

'''Materials'''

*Ketometer - Precision xtra ketone body / glucose monitor (Abbot)
*Ketone Body Test Strips
*Scale
*Bowl for weighing mice
*echoMRI if the mice differ in body fat levels (see below)
*Syringes
*1.5 mg/mL bOHB in PBS (make as 15mg bOHB in 10mL of PBS, sterile filtered). This will correspond to 1 U/kg injections. If you are using a higher or lower dose of bOHB, add more or less to the 10 mL of PBS, so that injections are 10 uL/g of mass.
*This may need to be adjusted depending on the ketone sensitivity of the mice, and this is based on PGC-1a on chow.
*In general you want the bOHB to increase blood bOHB levels by about 10 to 12-fold (bringing bOHB levels form around .5mmol/L up to the 5 to 6 mmol/L range in the most responsive of your two groups. If your response is lower or higher, you will probably have to change your dose and retry.
*Timer

'''Protocol'''

*If testing fasted ketone tolerance: Remove food from mice for about 6h by putting them in a fresh cage. Add “do not feed” acetate to cages, and ideally move cage to procedure room. Try to make sure that the mice are in a quiet, undisturbed temperature controlled room with the lights on. Typically starve the mice at 8AM and aim to start injections at 2PM
*Prepare a 1 g/10mL solution of glucose in the unlikely case that some animals become hyperketonemic.
*Weigh/MRI mice ahead of time, mark tails if necessary with different colors for rapid identification and take fasting ketone measurement via a tail clip.
*Prepare bOHB syringes with 1.5 g/KG mouse weight (ie for a 30g mouse, 300 uL).
*At approximately 1 min intervals, inject appropriate amount of bOHB into intraperitoneal cavity of the mouse. Immobilize mouse and restrain tail with one hand. Aim needle for peritoneal space, between the midline and the hip bone. Insert syringe (do not inject) into cavity. Eject syringe.
*At 15 minute intervals (normally 15, 30, 45, 60, 75, & 90 min), take blood bOHB measurements from tail vein. If needed re-snip the tail tip to expose the vein. When measuring BOHB just lift the tail of the mouse, while leaving it in the cage, rather than removing and restraining the mouse which can be more stressful.
*Analyze data by both % change from fasting ketones and absolute values. Our preferred outcome is to report fasting ketone levels and report percent change as a figure.
*The preferred statistical model is a mixed linear model using the time points as ordinate values and testing for a main effect or an interaction of the treatment/genotype. Use the R lme4 package for this.

First Strand cDNA Synthesis (AB Kit)

2019-07-24T15:16:23Z

Snyderds: adjusted quantity of RNA added

[[Category:qPCR]]
[[Category:RNA]]
[[Category:Transcription]]
[[Category:mRNA]]
[[Category:Mouse Work]]

==Materials==
* Purified RNA
* High Capacity cDNA Reverse Transcription Kit without RNAse inhibitor (AB 4368813 for 1000 reactions, see http://products.invitrogen.com/ivgn/product/4368813)

==Protocol==
* Generate Master Mix Containing:

{| border="1"
|+ Master Mix Components
! scope="col" | Component
! scope="col" | Per Reaction
! scope="col" | Per 10 Reactions
|-
! scope="row" | 10X RT Buffer
| 2 || 20
|-
! scope="row" | 25X dNTP Mix (100 mM)
| 0.8 || 8
|-
! scope="row" | 10X RT Random Primers
| 2 || 20
|-
! scope="row" | MultiScribe Reverse Transcriptase
| 1 || 10
|-
! scope="row" | Nuclease Free Water
| 4.2 || 42
|-
|}

* Add 10 uL Master mix per well into tube or plate
* Add 10 ug RNA in a final volume of 20 uL to each well. Since results will be normalized in the end, standardizing RNA quantities at this step is unnecessary. Total volume can be adjusted down if RNA is in short supply.
* Run the PCR Program '''AB Reaction Protocol'''. Takes ~2h:
** 10 min at 25C
** 120 min at 37C
** 5 min at 85C

QPCR

2019-02-19T20:25:51Z

Snyderds: added link to google calendar

__NOTOC__
==Real Time qPCR==
===Materials===
*cDNA: see [[First_Strand_cDNA_Synthesis_(AB_Kit)|First Strand cDNA Synthesis (AB Kit)]] for details
*SYBR Green PCR Master Mix Applied Biosystems (ThermoFisher Catalog # 4367659; [https://www.thermofisher.com/order/catalog/product/4367659 Vendor Link])
*384 well qPCR plate (ThermoFisher Catalog # 4309849) and covers (Catalog # 4360954)
*Primers (Dilute to 0.4 uM mixture of fwd and rev. From 100 uM stocks- the 100uM stock is prepared by adding 227 uL of distilled water to 22.7nmol of a gene as an example- this is 4uL Forward Primer, 4 uL Reverse Primer and 992 uL Water). This primer mix can be stored at -20 in a Working Primers box. Design primers according to [[Primer Design for qPCR]]

===Plate Preparation===
#Book 2h on qPCR machine by signing up on the sheet in room 7013 and on the [https://calendar.google.com/calendar/embed?src=umich.edu_jkmcg58uidmngdh0sutt1d36og%40group.calendar.google.com&ctz=America%2FDetroit google calendar]
#Prepare cDNA and dilute in water in a 96 well plate. Typically a 20x dilution of cDNA leaves enough to be detected.
#Get optically clear 384 well plate and keep on paper towel. Do not touch bottom of plate.
#Sketch out the plate in your notes. Typically rows are different primers while columns are different cDNA's
#Calculate how many samples x how many replicates/per sample (start with 3 or 4 until you are consistent enough technically to decrease). This will be the number of wells need for each primer.
#Prepare a Primer/SYBR Green mixture for each primer. For each well you will need 5 uL SYBR green + 2.5 uL Primer working stock, so if you have calculated you need 10 wells per primer that is 50 uL SYBR Green + 25 uL Primers. Make up 10-20% more than you need.
#Using the repeater multichannel pipette put on 2 or 3 tips (depending on your plate arrangement) and set to aspirate however many samples you have and dispense 7.5 uL per well.
#Dispense 7.5 uL of Primer/SYBR mixture into each well, dispensing at the bottom of the well.
#Using the ClipTip multichannel add 2.5 uL of cDNA to each applicable well. You don't need to change tips between wells.
#Once the plate is completed, carefully put an optically clear cover on it using the plastic square to ensure the edges are sealed, being very careful not to leave fingerprints on the seal.
#You can prepare the plate ~3h beforehand, keeping it at 4C until the machine is ready.
#Immediately before the run spin the plate briefly (2 mins at 4000 RPM) in the swinging bucket centrifuge.

===Run Protocol===
* Set up run using [[Set up qPCR Run on Thermo Cloud and QuantStudio|Thermo Cloud/QuantStudio]] or [[Set up qPCR Run on Roche LightCycler|Roche LightCycler]]

==Calculations==
see http://www.ncbi.nlm.nih.gov/pmc/articles/PMC55695 for considerations on calculations

[[ Category: Molecular Biology ]]
[[ Category: RNA ]]
[[ Category: qPCR ]]
[[ Category: Transcription ]]
[[ Category: Expression ]]

User:Snyderds

2019-01-22T18:07:20Z

Snyderds: Created page with " Category: SOP Category: Lab Safety '''Laboratory Director (LD) Approval is Required Prior to Performing this Procedure ''' ==Description== This standard operating..."

[[ 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 corrosive chemicals. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling corrosive chemicals.

A corrosive chemical is defined by meeting one the following:
*Corrosive chemicals will damage substances that they make contact with and they are immediately dangerous to living tissue.
*Most corrosive chemicals are acids and bases however; other chemicals can be corrosive as well. Always check chemical product information.
*Some common corrosive acids include hydrochloric acid, sulfuric acid, and nitric acid. Examples of corrosive bases are ammonium hydroxide, potassium hydroxide, and sodium hydroxide.
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.

==Process==
Refer to the steps in the protocol and the Safety Data Sheet (SDS) for using the corrosive chemical in question for information regarding the steps needed to securely handle that particular corrosive chemical.

==Potential Hazards==
Corrosive chemicals can cause destruction of living tissue by chemical action at the site of contact and can be solids, liquids, or gases. Corrosive effects not only can occur on the skin and eyes, but also in the respiratory tract and, in the case of ingestion, in the gastrointestinal tract as well. While acids and bases are familiar corrosives, many other materials are corrosive to the body as well (including [[SOP - Bleach]] and [[SOP- Phenol]]).

Review the Safety Data Sheet (SDS) of the material to determine if it is corrosive and to identify other hazards. If the material also has other physical hazards or health hazards, follow those appropriate SOP as well.

Refer to SOP templates for hydrofluoric [http://ehs.umich.edu/wp-content/uploads/sites/37/2016/02/HydrofluoricAcid.docx|hydrofluoric acid], perchloric [http://ehs.umich.edu/wp-content/uploads/sites/37/2016/02/PerchloricAcid.docx|perchloric acid], [[SOP- Phenol]], and [[SOP - Bleach]].

Acutely toxic and reactive acids meeting the MIOSHA definition of a Particularly Hazardous Substance (see below) require customized SOPs with laboratory director approval that incorporate their toxic and/or reactive hazards.

'''Notes'''

The University of [http://ehs.umich.edu/wp-content/uploads/sites/37/2016/03/ChemicalHygienePlan.pdf|Michigan’s Chemical Hygiene Plan] requires specific measures to take in order to ensure laboratory employee protection, including provisions for additional employee protection for work with Particularly Hazardous Substances. These include "select carcinogens," reproductive toxins and substances which have a [http://www.osha.gov/pls/oshaweb/owadisp.show_document?p_table=INTERPRETATIONS&p_id=20048|high degree of acute toxicity], e.g., hydrogen cyanide, hydrogen sulfide, and nitrogen dioxide.
Specific consideration shall be given to the following provisions which shall be included where appropriate for each '''Particularly Hazardous Substance''':
* Establishment of a designated area;
* Use of containment devices such as fume hoods or glove boxes;
* Procedures for safe removal of contaminated waste; and
* Decontamination procedures.
Environment, Health & Safety (EHS) can assist researchers by providing additional information on working with Particularly Hazardous Substances. General guidelines and recommendations for the safe handling, use and control of hazardous chemicals and particularly hazardous substances can be found in a chemical’s SDS [http://ehs.umich.edu/research-clinical/chemical/safety-data-sheets/|SDS] and in other [http://ehs.umich.edu/research-clinical/chemical/safety-data-sheets/|chemical hazard references].

==Engineering Controls==
*Where the eyes or body of any person may be exposed to corrosive chemicals, 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.
*A safety or drench shower should be available in a nearby location where the corrosive material is used.
*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.
*Manipulation of some corrosive materials 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 corrosive materials in the laboratory and are the preferred ventilation control device.

==Work Practice Controls==
*Segregate the various types of corrosives. Separate acids and bases. Liquids and solids should also be separated. Specially designed corrosion resistant cabinets should be used for the storage of large quantities of corrosive materials. Store corrosives on plastic trays. Do not store corrosive materials on high cabinets or shelves.
*Nitric acid should be stored in secondary containment in a well-ventilated area that is separated from organics and other combustible materials and incompatibles. Also refer to the SOP for hydrofluoric acid, nitric acid, picric acid and sulfuric acid.
*'''Containers''': All corrosive chemicals must be clearly labeled with the correct chemical name. Handwritten labels are acceptable; chemical formulas and structural formulas are not acceptable.

'''Notes'''
Employers may choose to label secondary, portable or in-house workplace containers with label alternatives that meet the requirements for the new Hazard Communication Standard (HCS). Alternative labeling systems such as the National Fire Protection Association (NFPA) the Hazardous Material Information System (HMIS) are permitted for workplace containers. However, the information supplied on these labels must be consistent with the revised HCS, e.g., no hazard warnings or pictograms that conflict with GHS.

==Personal Protective Equipment==
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 EHS representative.

Eye protection in the form of safety glasses must be worn at all times when handling corrosive materials. 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 ''American National Standard for Occupational and Educational Personal Eye and Face Protection Devices'' (ANSI/ISEA Z87.1) and must be equipped with side shields (that also meet the requirements of ANSI/ISEA Z87.1). 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. It is recommended that face shields be worn when a splash potential exists with corrosive materials.

Gloves must be worn when handling corrosive chemicals. Disposable nitrile gloves provide adequate protection against accidental hand contact with small quantities of most laboratory chemicals. Below are some glove material recommendations for some common acids.
{| class="wikitable"
|-
! '''Acid''' !! '''Recommended Glove Materials'''
|-
| Acetic Acid ||
*Butyl (Unsupported)
*Latex (Unsupported Natural Rubber)
*Neoprene
*Nitrile (Unsupported)
*Polyvinyl Chloride (PVC)
|-
| Hydrochloric Acid ||
*Butyl
*Neoprene
*Nitrile
*Polyvinyl Chloride (PVC)
|-
| Nitric Oxide ||
*Butyl
*Natural Rubber
*Neoprene
|-
| Phosphoric Acid ||
*Natural Rubber
*Neoprene
*Nitrile
*Polyvinyl Chloride (PVC)
|-
| Sulfuric Acid||
*Butyl
*Neoprene
*Polyethylene
*Polyvinyl Chloride (PVC)
|}
'''Note''' The recommendations above are based on typical laboratory concentrations and uses. Refer to the acid’s SDS as well as Glove Compatibility Charts for each particular concentration of acid for additional recommendations, especially for unusual concentrations or use of a particular acid.

Lab workers should contact EHS for advice on chemical resistant glove selection when direct or prolonged contact with hazardous chemicals is anticipated. The EHS [http://ehs.umich.edu/research-clinical/planning-safe-research/glove-compatibility-chart/|Glove Compatibility] webpage can also be used to determine the recommended gloves.

Lab coats, long pants, closed toed shoes and long sleeved clothing should be worn when handling corrosive materials. Additional protective clothing should be worn if the possibility of skin contact is likely.

Safety shielding is required any time there is a risk of explosion, splash hazard or a highly exothermic reaction. All manipulations of corrosive materials 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.

==Transportation and Storage==
*Transport corrosives 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 – many corrosive materials are incompatible with each other – oxidizing acids are incompatible with organic acids, and acids are incompatible with bases. *Review the chemical’s Safety Data Sheet (SDS) for additional incompatibility information.
Avoid storing on the floor. If storing on the floor is necessary, use secondary containment.

==Waste Disposal==
Most spent, unused and expired corrosive materials are considered hazardous wastes and must be collected and disposed of within 90-days by EHS’s Hazardous Materials Management (HMM) Group. Contact EHS-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also, refer to the [http://ehs.umich.edu/haz-waste/|EHS 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:
*'''Personnel''': Immediately flush contaminated area with copious amounts of water after contact with corrosive materials. Remove any clothing and/or jewelry to facilitate removal of chemicals. *If a delayed response is noted report immediately for medical attention. Be prepared to detail what chemicals were involved.
*If the incident involves '''Hydrofluoric acid (HF)''', seek immediate medical attention. Also, refer to the [http://ehs.umich.edu/wp-content/uploads/sites/37/2016/02/HydrofluoricAcid.docx|EHS HF SOP].
*If there is any doubt about the severity of the injury, seek immediate medical attention.
Contact EHS 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 [http://www.workconnections.umich.edu/employees/work-related-illness-injury/step-one/|Illness and Injury Report Form]. Follow the directions on the Work Connections website [http://www.workconnections.umich.edu/treatment.html|Where to go for treatment] to obtain proper medical treatment and follow-up.
*Complete the 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==
'''Area''': Decontamination procedures vary depending on the material being handled. The corrosivity of some materials can be neutralized with other reagents. Special neutralizing agents should be on hand to decontaminate areas.
*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.
For additional information regarding spill response procedures, refer to the EHS Hazardous Waste Spill Response Web page.

For additional information regarding spill response procedures, refer to the [http://ehs.umich.edu/hazardous-waste/spill-response/|EHS Hazardous Waste Spill Response Webpage].

==Additional Spill Response Steps:==

'''MINOR CHEMICAL SPILL'''
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. In the event of a minor chemical spill, use the following information for a safe cleanup process.
*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.
*Use proper personal protective equipment (PPE) as noted 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 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-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.
A '''major''' (large) chemical spill requires active assistance from emergency personnel. In the event of a major chemical spill, use the following information for a safe spill response process.
*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 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.

==Emergency Reporting==
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 [http://dpss.umich.edu/emergency-management/alert/|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 [http://ehs.umich.edu/education/|EHS’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)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 14:48, 13 June 2017 (UTC)
Laboratory Director - Dave Bridges Revision Date - 2019-01-22

Set up qPCR Run on Thermo Cloud and QuantStudio

2018-12-18T19:17:30Z

Snyderds: added link to "QPCR Analysis on ThermoCloud"

==Set Up Plate Layout==

* Log into Thermo Cloud ([https://apps.thermofisher.com/apps/dashboard/#/ https://apps.thermofisher.com/apps/dashboard] using the login information for your lab (the Bridges Lab account data is located on the side of the machine.))
*Click file folder icon (on the left side of the screen under the home icon), then '''New Folder''' (add your name), and '''Create'''.
*Click '''Create Project''', give your project a name, and assign it to your folder.
* Under '''My Apps''' on the right hand side click on '''Design and Analysis Application (qPCR)'''.
* Copy a plate layout file (click '''Plate''' and choose '''Export plate setup''' from the '''Action''' menu) and save it as a CSV file. (to create a template in the program, see addendum below)
** For each well you are using you need to enter the following:
*** Sample Name
*** Target Name
*** Task (Usually UNKNOWN)
*** Reporter (SYBR)
*** Quencher (NFQ-MGB)
** Leave the rest of the plate blank
* Click on '''New''' under '''Create new experiment'''.
* Leave the file name as the default (Date/ID)
** Change '''Block type''' to '''384-Well Block'''.
** Change '''Chemistry''' to '''SYBR Green Reagents'''.
** Click Next or on the '''Method''' tab and change volume to 10 uL (or leave at 20 uL if you are using a total volume of 20uL/well)
** Click Next or on the '''Plate''' tab. Click on '''Actions''' then '''Import plate setup''' and select your plate layout file. Check the visual representation of your plate layout and make sure it is correct.
** Click Next or on the '''Run''' tab. The software will save your plate layout and upload it to thermo cloud for access on the Quant Studio5
*After you plate runs, refer to [[QPCR Analysis on ThermoCloud|qPCR Analysis on ThermoCloud protocol]].

==Addendum==

*In the '''Plate''' Tab on the plate schematic, outline wells, type in sample number--> Enter to assign (repeat for all samples)-->Outline wells, type in target (primer)-->Enter to assign--> Next

[[ Category: Molecular Biology ]]
[[ Category: RNA ]]
[[ Category: qPCR ]]
[[ Category: Transcription ]]
[[ Category: Expression ]]
[[ Category: Equipment Protocols ]]

QPCR Analysis on ThermoCloud

2018-12-04T16:49:17Z

Snyderds:

*Log into Thermo Cloud ([https://apps.thermofisher.com/apps/dashboard/#/ https://apps.thermofisher.com/apps/dashboard] using the login information for your lab (the Bridges Lab account data is located on the side of the machine.)
*Click on the '''File Folder icon''' and find your folder (or the folder in which you saved your run).
*Select the file with extension '''.eds'''. The program will automatically take you to the '''Results''' tab of your run.
*Above the graph, use the dropdown box to select '''Melt Curve'''.
*Click on '''Target''' above the plate schematic to select one target at a time.
**Verify that there is only one (prominent) peak for each of your targets.
**Verify that each of your replicates amplified for each target.
**If there is a target that yields 2+ peaks over all samples, mark that target as unusable in the '''Primer Database'''.
**Exclude samples that did not pass the above criteria by clicking the well (not the plate schematic) and select '''Omit''' under the '''Action''' dropdown box above the plate schematic.
*Check the technical replicates by choosing the '''Table View''' and arranging them according to target. Verify that the '''CT''' values have <1.5 difference. Omit outliers.
*Click '''Next''' and '''Export''' your results to the appropriate project folder.
*Under '''App Connect''' on the right hand side click on '''Relative Quantification''' (qPCR). Open your run.
*Under the '''Overview''' tab, assign biological groups to each of your samples (if needed).
*If you need to omit samples from the analysis, do so under the '''Data Review''' tab by clicking on the affected target, highlighting, and omitting the wells with outliers.
*Under the '''Analysis''' tab click the blue '''Analyze''' button in the top right corner to visualize your adjusted data.
*'''Export''' your data to the appropriate location.

[[ Category: Molecular Biology ]]
[[ Category: RNA ]]
[[ Category: qPCR ]]
[[ Category: Transcription ]]
[[ Category: Expression ]]
[[ Category: Equipment Protocols ]]

Seahorse - Mitochondrial Stress Test

2017-06-14T17:02:55Z

Snyderds: /* Protocol */ added "remove lid" in 2nd to last step

[[ Category: Seahorse ]]
[[ Category: Mitochondrial Function ]]

==Materials==
* [[Seahorse XF Media]]
* XF Cartridge
* Cells in XF24 plate, either grown in the plate or seeded at a previously established density where the OCR is 40-500 mL/min.
* Injection stock solutions (1 mM Oligomycin, 1 mM FCCP, 1 mM Rotenone and 1 mM Antimycin A) made up in DMSO at 1000X. Aliquots in the -20 in '''Seahorse Reagents''' box.
* Check if the machine is free.

==Protocol==

===The Day Before the Assay===
* Hydrate a cartridge by placing 1 mL/well of XF Calibration Media into each well of a XF24 cartridge. Place in non-CO2, humidified, 37C incubator overnight to hydrate.
* Ensure that there is sufficient XF media and stock solutions.
* Prepare cells if necessary, leave in CO2 incubator overnight. Make sure to leave blank wells (typically A5, B3, C4, D2).

===The Day of the Assay===
* Prepare media by the following steps:
** Place 100mL in non-CO2, humidified, 37C incubator for ~30 minutes to warm up.
** Add the following solutions (these are standard concentrations and may be slightly different for your cell line of interest):
*** 10 mM Glucose (1 mL of a 1M stock into 100 mL)
*** 2 mM Glutamine (1 mL of a 200 mM stock)
*** 1 mM Pyruvate (100 uL of a 1M stock)
** Re-adjust the pH to 7.4 (7.35-7.45)
* Wash cells twice with 500 uL of pre-warmed media, add 500 uL final volume to each well
* Incubate for 1h in non-CO2, humidified, 37C incubator to get rid of any extra CO2
* While plate is incubating, prepare injection solutions in 1.8 mL of prepared XF Media. These are standard concentrations, and should be optimized for your system (especially FCCP)
** '''Tube A''' 18 uL Oligomycin
** '''Tube B''' 18 uL FCCP
** '''Tube C''' 9 uL Rotenone and 9 uL Antimycin A
* Add the compounds to the injection ports in the cartridge (A, bottom right; B bottom left; C top right). '''Make sure the barcode is on the right hand side.'''
* Set up your protocol, editing a similar template and saving with the date and experiment type
* Start the protocol, inserting the cartridge with the lid removed. '''Make sure the barcode is on the right hand side.'''
* Once the cartridge is calibrated insert the cell plate at the prompt. If the calibration fails, abort the run and try the calibration again

Seahorse - Mitochondrial Stress Test

2017-06-14T13:39:11Z

Snyderds:

[[ Category: Seahorse ]]
[[ Category: Mitochondrial Function ]]

==Materials==
* [[Seahorse XF Media]]
* XF Cartridge
* Cells in XF24 plate, either grown in the plate or seeded at a previously established density where the OCR is 40-500 mL/min.
* Injection stock solutions (1 mM Oligomycin, 1 mM FCCP, 1 mM Rotenone and 1 mM Antimycin A) made up in DMSO at 1000X. Aliquots in the -20 in '''Seahorse Reagents''' box.
* Check if the machine is free.

==Protocol==

===The Day Before the Assay===
* Hydrate a cartridge by placing 1 mL/well of XF Calibration Media into each well of a XF24 cartridge. Place in non-CO2, humidified, 37C incubator overnight to hydrate.
* Ensure that there is sufficient XF media and stock solutions.
* Prepare cells if necessary, leave in CO2 incubator overnight. Make sure to leave blank wells (typically A5, B3, C4, D2).

===The Day of the Assay===
* Prepare media by the following steps:
** Place 100mL in non-CO2, humidified, 37C incubator for ~30 minutes to warm up.
** Add the following solutions (these are standard concentrations and may be slightly different for your cell line of interest):
*** 10 mM Glucose (1 mL of a 1M stock into 100 mL)
*** 2 mM Glutamine (1 mL of a 200 mM stock)
*** 1 mM Pyruvate (100 uL of a 1M stock)
** Re-adjust the pH to 7.4 (7.35-7.45)
* Wash cells twice with 500 uL of pre-warmed media, add 500 uL final volume to each well
* Incubate for 1h in non-CO2, humidified, 37C incubator to get rid of any extra CO2
* While plate is incubating, prepare injection solutions in 1.8 mL of prepared XF Media. These are standard concentrations, and should be optimized for your system (especially FCCP)
** '''Tube A''' 18 uL Oligomycin
** '''Tube B''' 18 uL FCCP
** '''Tube C''' 9 uL Rotenone and 9 uL Antimycin A
* Add the compounds to the injection ports in the cartridge (A, bottom right; B bottom left; C top right). '''Make sure the barcode is on the right hand side.'''
* Set up your protocol, editing a similar template and saving with the date and experiment type
* Start the protocol, inserting the cartridge. '''Make sure the barcode is on the right hand side.'''
* Once the cartridge is calibrated insert the cell plate at the prompt. If the calibration fails, abort the run and try the calibration again

FDB Muscle Dissection from Mice

2017-06-13T19:46:12Z

Snyderds:

This protocol is modified from section 3.1.2 of [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3523695/ Isolation and Culture of Skeletal Muscle Myofibers as a Means to Analyze Satellite Cells] and the first part of the following video with accompanying protocol [https://www.jove.com/video/53551/dna-electroporation-isolation-and-imaging-of-myofibers DNA Electroporation, Isolation and Imaging of Myofibers | Protocol].

<big>'''Materials'''</big>
* Tweezers
* Standard operating scissors
* Surgical scissors
* Forceps
* Dissection stage with 7 pins
* Anesthetic

<big>'''Flexor Digitorum Brevis Muscle Extraction'''</big>

* Sacrifice the animal with anesthetic gas inhalation followed by cervical dislocation to ensure death.
* Remove the hind feet above the ankle joint using standard operating scissors.
* Pin one foot tight, sole facing up, onto the dissection stage with pins through each of the 5 toe tips and one through the plantar side of the ankle.
* Starting at the ankle, snip the skin straight up the side of the foot along the hairline towards the toes, being careful not to nick the muscle beneath the skin. Repeat on the other side of foot.
* Holding the skin at the base of the ankle with forceps, cut the skin across the heel.
* Lift the skin flap with forceps. With your scissors pointed toward the skin, parallel with the tendons so as to not nick the muscle, snip the underlying connective tissue, effectively removing the skin.
* Continue snipping connective tissue and gently pulling up on the skin until the tendons of the digits are exposed. Pin the skin onto the stage above the foot.
* To isolate the tendons, insert tweezers between the bone and the 5 tendons below where the muscle becomes thicker. Leave enough room to grasp the bundle of tendons with the forceps below the tweezers.
* Grasp the tendon bundle at the base, below the tweezers, ensuring that the muscle tissue is not damaged. Remove tweezers. To remove the FDB muscle bundle, cut the large tendon at the heel to free the tendon from the bone. Holding the tendon with forceps, dissect the FDB bundle from the large white tendon, carefully removing any connective tissue attached to the FDB with scissors. If done properly the bundle will easily lift off of the underlying tendons revealing bright white tendons leading to the individual digits. Cut the FDB at the tendons near the toes freeing the bundle from the foot. Err on the side of retaining tendon tissue on the dissected muscle and avoid damaging the FDB muscle.
* Place the entire muscle bundle in the well containing KRBH.
* Repeat on the contralateral foot and place in the same well.

FDB Muscle Dissection from Mice

2017-06-13T19:08:28Z

Snyderds:

Fdb muscle dissection from mice

2017-06-13T17:52:44Z

Snyderds: Snyderds moved page Fdb muscle dissection from mice to FDB Muscle Dissection from Mice

#REDIRECT [[FDB Muscle Dissection from Mice]]

FDB Muscle Dissection from Mice

2017-06-13T17:52:44Z

Snyderds: Snyderds moved page Fdb muscle dissection from mice to FDB Muscle Dissection from Mice

This protocol is modified from section 3.2.1 of [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3523695/ Isolation and Culture of Skeletal Muscle Myofibers as a Means to Analyze Satellite Cells] and the first part of the following video with accompanying protocol [https://www.jove.com/video/53551/dna-electroporation-isolation-and-imaging-of-myofibers DNA Electroporation, Isolation and Imaging of Myofibers | Protocol]

<big>'''Materials'''</big>
* Bulleted list item

FDB Muscle Dissection from Mice

2017-06-13T17:11:31Z

Snyderds: Created page with "This protocol is modified from section 3.2.1 of [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3523695/ Isolation and Culture of Skeletal Muscle Myofibers as a Means to Analyze..."

Isolation of Single Fibers for Seahorse Assays

2017-06-13T17:06:35Z

Snyderds:

This protocol is modified from https://www.agilent.com/cs/library/applications/5991-7148EN.pdf

==Materials==
* Seahorse Plate (XF24) and Cartridge
* Matrigel, slowly thawed and aliquoted into 100 uL aliquots. Stored at -20
* [[KRBH Buffer]], about 20 mL pre-warmed to 37C
* Collagenase
* DMEM with no Phenol (Invitrogen cat# [[https://www.thermofisher.com/order/catalog/product/21063029 21063-029]]), need about 20 mL
** Prepare by adding 2% FBS (400 uL) and 200 uL PSG to a 50 mL conical tube
* Assay Media - [[aCSF Media]], about 10 mL pre-warmed to 37C

==Muscle Fiber Dissection and Digestion==
* Place ~20 mL KRBH, ~20 mL DMEM and ~100 mL aCSF Media in a 37C water bath to warm.
* Sterilize surgical tools for ~30 min and move mice to the procedure room.
* Dissect FDB muscles from mice (see [[FDB Muscle Dissection from Mice]]). Place both muscles from a mouse in a single 12-well containing 1 mL of pre-warmed KRBH.
* Weigh out enough collagenase to make 1 mL of a 4 mg/mL solution for each mouse. Add DMEM to make the 4 mg/mL solution
* Gently transfer muscles to wells containing 1 mL of the collagenase solution.
* Incubate at 37C for 1.5-2h to allow for digestion.
* Gently transfer muscles to a new well containing 1 mL DMEM.
* Pipet muscles up and down gently 5-10 times until the muscle is dissociated. If dissociation did not yield a sufficient number of fibers, digest for a further 15-30 minutes and repeat the process.
* Try to pick out any obvious undigested material such as tendons using sterile tweezers.
* Prepare XF24 plate by combining 100 uL matrigel with 100 uL media. Mix well by tapping.
* Add 3 uL of this mixture to each well of a XF24 plate. Disperse the matrigel by hitting the plate against your hand to cover the surface.
* Leave uncovered in the tissue culture hood for 30 minutes for the matrigel to dry.
* Gently swirl the digested fibers and pipet 50 uL of the digested fiber mixture to each well.
* Fibers should attach within 5 minutes. The assay can now be run, or cells can incubate overnight prior to the assay.

==Mitochondrial Stress Test==

Spill Response Procedures for Infectious agents and Recombinant DNA

2017-06-07T19:58:38Z

Snyderds:

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)

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 - Vacuum Pumps

2017-06-07T19:56:12Z

Snyderds:

[[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)

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

SOP - Sensitizers

2017-06-07T19:53:48Z

Snyderds:

[[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)

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 - Irritants

2017-06-07T19:52:46Z

Snyderds:

[[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)

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 - Compressed Gases

2017-06-07T19:44:48Z

Snyderds:

[[ 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)

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

SOP - Autoclave

2017-06-07T19:43:09Z

Snyderds:

[[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)

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 - Rapamycin

2017-06-07T19:37:28Z

Snyderds:

[[ 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)

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 - Needle Capping

2017-06-07T19:33:20Z

Snyderds:

[[ 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)

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 - Formalin and Paraformaldehyde

2017-06-07T19:31:35Z

Snyderds:

[[ 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)

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

SOP - Flammables and Combustibles

2017-06-07T19:15:09Z

Snyderds:

[[ 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)

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 - Electrophoresis

2017-06-07T19:12:24Z

Snyderds:

[[ 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)

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 - Chloroform

2017-06-07T18:50:36Z

Snyderds:

[[ 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)

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 - Centrifuge Safety

2017-06-07T18:49:30Z

Snyderds:

[[ 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)

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 - Carcinogens

2017-06-07T18:48:58Z

Snyderds:

[[ 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)

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

SOP - Bleach

2017-06-07T18:47:42Z

Snyderds:

[[ 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)

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 - Animal Anesthetics

2017-06-07T18:34:34Z

Snyderds:

[[ 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)
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