Bridges Lab Protocols - User contributions [en]

High Fat Diet Treatment

2018-11-28T18:42:40Z

Cdgleason: update diet table

[[ Category: Diet ]]
[[ Category: Animal Studies ]]
[[ Category: Glucose Homeostasis ]]
[[ Category: Metabolism ]]
[[ Category: Mouse Work ]]

To place mice on a high fat diet, here is the general protocol:

For C57BL/6J mice, order so that they are 9 weeks of age on the monday of delivery. Start the diet on the Wednesday (10 days later).

==Diet Choices==

For more information about diets see [http://www.researchdiets.com/system/resources/BAhbBlsHOgZmIisyMDEyLzA0LzIwLzE0XzM5XzQwXzMxN19ESU9fU2VyaWVzLnBkZg/DIO%20Series.pdf research diets]
for caloric content in future diets, use metabolizable energy, not physiological fuel value. Metabolizable energy is more accurate as it is derived from bomb calorimetry

{| border=1
|+ Food Souces and Notes about their Composition
|-
! Food
! Vendor
! Code
! Caloric Content
! Fat
! Protein
! Sucrose
! Starch
! Notes
|-
| Normal Chow Diet
| Lab Diet
| 5L0D
| 2.91 kcal/g
| 5%
| 24%
| 3.7%
| 32%
| Provided by ULAM
|-
| High Fat Diet
| Research Diets
| D12451
| 4.73 kcal/g
| 45%
| 20%
| 17%
| 7%
| Pink
|-
| Control Diet (High Sucrose)
| Research Diets
| D12450B
| 3.85 kcal/g
| 10%
| 20%
| 35%
| 31%
| Yellow
|-
| Control Diet (High Starch)
| Research Diets
| D12450H
| 3.85 kcal/g
| 10%
| 20%
| 17%
| 44%
| Peach
|-
| Control Diet (KD)
| Research Diets
| D1053001
| 3.8 kcal/g
| 10%
| 15%
| 0%
| 75%
| Pellet form
|-
| Ketogenic Diet
| Research Diets
| D17053002
| 6.4 kcal/g
| 85%
| 15%
| 0%
| 0%
| Paste form
|}

==Treatment Protocol==

# Start when mice are 10 weeks of age.
# For High Fat Diet mice place in cages 4 mice/cage as early as possible. Can us 5 mice/cage for normal chow/control diets.
# Re-cage (and ear tag mice if necessary) mice and replace food with ~400g food in the container. Record this weight.
# This food should last 2 weeks.
# Weigh mice at ~10 AM either weekly or bi-weekly and re-weigh food to determine food intake. Top up to 400g.
# Generally treatment lasts 12 weeks.

SOP - Phenol

2018-10-04T03:55:27Z

Cdgleason: certification of above SOP

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

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

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

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

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

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

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

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

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

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

• Keep containers closed as much as possible.

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

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

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

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

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

• Keep container in cool, well-ventilated area.

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

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

• Store below eye level.

• Avoid storing on the floor.

• Avoid ignition sources.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

SOP - Chloroform

2018-10-04T03:54:30Z

Cdgleason: certification of above SOP

[[ Category: SOP ]]

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

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

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

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

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

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

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

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

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

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

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

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

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

==Exposures/Unintended Contact==

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

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

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

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

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

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

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

===Spill Response Steps:===

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

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

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

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

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

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

* [[User:Davebridges|Davebridges]] ([[User talk:Davebridges|talk]]) 14:42, 15 August 2016 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 16:15, 31 October 2016 (UTC)
*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:09, 5 June 2017 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 18:50, 7 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:01, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:06, 12 June 2017 (UTC)
*[[User:Cdgleason|Cdgleason]] ([[User talk:Cdgleason|talk]]) 03:54, 4 October 2018 (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