Slides adapted from: Mr. Wayne Wood, Manager ,

1. Essentials of Laboratory Safety Heather D. Durham, PhD Chair, Laboratory Safety Committee Montreal Neurological Institute Slides adapted from: Mr. Wayne Wood, Manager , Environmental Health and Safety, McGill University and Howard Hughes Medical Institute, Online Safety Course http://www.practicingsafescience.org/

2. MODULE 1: General Laboratory Safety Getting Ready to Work Working in the Lab Emergency Response Individual Protection Chemical Hoods Biosafety Cabinets Tissue Culture Using Autoclaves Chemical Storage Leaving the Lab MODULE 2: WHMIS Classification of Products Main Elements of WHMIS Labels MSDS Organization of the Presentation You are expected to be familiar with the information on the website of McGill Environmental Health and Safety (EHS) http://www.mcgill.ca/ehs/ Additional resources are listed on the website for the MNI Laboratory Safety Committeehttp://www.mni.mcgill.ca/

3. MODULE 1: GENERAL LABORATORY SAFETY All personnel must be aware of general occupational health and safety issues and procedures, have read the safety manuals applicable to their work, and completed the required training. Laboratory Safety Manual http://www.mcgill.ca/ehs/laboratory/labsafety/ Biosafety Manual http://www.mcgill.ca/ehs/laboratory/biosafety/ Radiation Safety Manual http://www.mcgill.ca/ehs/radiation/manual/ Consult your supervisor to determine the training required in your laboratory.

4. Before Lab Work, Get to Know: • Hazards of materials & agents and their prescribed safety procedures • Proper and safe use of all equipment • First get ready, then get to work!

5. Also Get to Know: • Emergency spill procedures, use of adsorbents and disinfectants • General Emergency Procedures for your Institution • Designated escape route and alternate • Location of fire extinguishers, eye wash, shower, first aid, and spill kits

6. Before Lab Work, Get to Know: • Emergency telephone numbers and reporting procedures For all emergencies call: 55-555

7. While Working in the Lab: • Authorized persons only • Identify EVERYTHING! • No food, beverages, tobacco products, or application of cosmetics • Follow Good Laboratory Practices • Practice Safe Science

8. Weekly/Monthly Lab Checks Download file: MNI laboratory safety inspection checklist.xls from MNI website • Fume Hoods/Biosafety Cabinets • Tubing, pressurized connections • Chemical storage • Cleanliness & Orderliness • Eye wash (purge) • Fire extinguisher • First Aid Kits

9. Evaluating Lab Hazards • Regular review of the types of hazards: • chemical • physical • biological • ergonomic • mechanical Minor Variation in Procedure

10. Causes of injury in laboratories

11. While Working in the Lab: • Report all: • Accidents • Injuries • Fires • Spills • Close calls • Malfunctioning equipment • Safety hazards 1 Disabling injury 10 Minor injuries 30 Property damage incidents 600 Close Calls

12. Emergency Response When you take a job in a new laboratory, one of the first things you should do is ask your supervisor to review with you the emergency response plans for the lab. Make particular note of the locations of: • Emergency telephone numbers. • Eyewash fountains and emergency showers. • Spill kits. • Fire extinguishers. • Emergency exits and evacuation routes.

13. Fire • Fire is the most potentially devastating emergency in the modern biology laboratory. It is imperative that you know how to prevent fires and be prepared to respond should a fire occur. • Preventing fires. Use of flammable solvents is a primary cause of lab fires. Always follow these prudent practices: • Use the smallest quantities of flammable solvents as practical. • Store stock quantities in flammables storage cabinets. • Separate flammable solvents from sources of ignition.Never use a Bunsen burner in any area where flammable solvents are handled.

14. Emergency Response to Fire • Know the fire emergency response procedures of your laboratory and institution procedures of your laboratory. • Keep the lab aisles and evacuation routes free of equipment and other objects that could obstruct safe passage. Remember: Safe passage is for your benefit and for the benefit of emergency responders. • Periodically practice emergency response procedures. • Follow these immediate procedures in case of a major lab fire: • Alert people to evacuate the area. • Activate nearest fire alarm or call for emergency response. • Assess the situation: Only if it is safe to do so, try to extinguish the fire. • Close lab doors to confine fire. • Have a person who is knowledgeable about the lab and the incident ready to assist emergency personnel.

15. While Working in the Lab:Individual Protection • Shoes with full coverage and good grip soles (no sandals) • Restrain long hair, loose clothing and jewelry • Wear a lab coat • Use appropriate eye, skin, and hand protection • Wash hands often

16. Howard Hughes Medical Institute: Online Safety Course http://www.practicingsafescience.org/ I'm here to help you load that TFA into the centrifuge. Oops! I forgot my eye protection. I have to go get it. • Well RUN for Pete's sake. Time is money!  • Eye protection? You don't need any eye protection.  • Okay. Take your time and be aware of your surroundings. It'd be ironic if you were to cause an accident by trying to avoid an accident, wouldn't it? Think ahead and plan your moves. Be careful of your surroundings.

17. Eye Protection • Protects against risk of flying objects or dust particles, splashes of hazardous materials or harmful rays

18. Safety Glasses • Unbreakable lenses of plastic or tempered glass • For light-to-moderate work • Can be prescription lenses • Do not interfere with contact lenses

19. Goggles • Work with significant risk of splash of chemicals or projectiles • Can be worn over prescription glasses

20. Face Shield • Work with significant risk of splash on face or possible explosion (including removing vials from liquid nitrogen) • Face shield protects face adequately but not eyes

21. Hand Protection • Protects against risk of cuts, abrasions, burns, or exposure to hazardous materials. • Requires selection of the appropriate chemical resistant gloves • Change gloves often – none are impermeable! • Wash your hands often to reduce contamination of yourself and others. • Do not touch door knobs, telephones, etc. with gloves – remove them. • If you must transfer your experiment to another room • put all materials, including clean gloves, in a tray with spill-proof edges • remove gloves and wash hands • Re-glove once you are in the alternate location

22. While Working in the Lab: • Follow universal precautions • Handle unknowns as if they were hazardous

23. While Working in the Lab: • Handle volatiles, aerosols or fine powders in a chemical fume hood Contain bioaerosols in a biological safety cabinet

24. Laboratory Chemical Hoods • The laboratory chemical hood is a ventilated enclosure that protects you from being exposed to chemical fumes, gases, and aerosols that are generated within the enclosure. Protection is provided by room air that is drawn into the hood and vented to the atmosphere. The hood ventilation provides further protection by diluting the concentration of flammable gases below explosion limits. • The sash must be lowered to the operating position or vapours will spill out. • The hood should always be on. Notify the facility staff or the safety office immediately if the hood is off or you observe contaminants escaping from it.

25. Using Laboratory Chemical Hoods • Plan your experiment. First, assess the risks of your experiment. If a laboratory chemical hood is required, place everything you need in the hood before starting. • Lower the sash. Always make sure the sash is pulled down to the marked level. The best protection is provided when the sash is brought to the lowest level for convenient operation. • Watch your activity. Use slow and deliberate motions. Place supplies so that they do not obstruct the airflow at the airfoil sill or the exhaust slots at the back of the hood. Do not work within four inches of the airfoil sill. • Keep the work area uncluttered. Do not store chemicals in the cabinet.

26. Storage of Hazardous Chemicals • Laboratory directors, managers and personnel are responsible for understanding the general principles governing storage of the various classes of chemicals and for complying with McGill’s policy. This includes segregation of incompatible chemicals, proper storage conditions (e.g., safety cabinets) and limits on volumes of certain chemicals that may be stored in the laboratory (e.g., flammable liquids). • See http://www.mcgill.ca/ehs/laboratory/labsafety/#3.6.3.1 • Section 4. Handling and Storage of Laboratory Chemicals; • Section 7. Laboratory Ventilation And Fume Hoods • Appendix 1: Flammability Classification (NFPA) and Permissible Container Sizes (OSHA)

27. Chemical Storage • Storage areas for corrosive, toxic, flammable, and highly reactive chemicals should be near a laboratory chemical hood. • Transfer hazardous chemicals from stock containers to use containers only in a laboratory chemical hood. • You should not store hazardous chemicals inside the hood.

28. Rules of Thumb for Safe Storage • Do not store more chemicals than you will need over a reasonable time • Always read the chemical's label and mark it with the date of receipt before storing. • Never store highly reactive chemicals for longer than 6 months. • Never store a chemical with an obscured or missing label. • Separate chemicals into compatible groups and store alphabetically within the groups (see section 4 of lab safety manual). • Designate separate storage areas for highly toxic chemicals. • Flammables, organic acids and organic bases in separate trays in a flammables’ cabinet • Inorganic acids, strong bases in separate trays in the cabinet under the fume hood • Toxic chemicals separately. • Oxidizers separately. • Corrosives separately. • Dry chemicals on shelves with raised edges, separated according to incompatibility. • Flammables should never be stored in a frost-free refrigerator/freezer.

29. Class II Biosafety Cabinets • The type of hood present in most tissue culture labs. • Protects you from being exposed to infectious aerosols that may be generated within the cabinet. You are protected by room air that is drawn into the cabinet front grill and the cabinet downward airflow. This combination of airflows quickly prevents respirable size particles from escaping into your lab. • Protects your cultures from microbialcontaminants that are ubiquitous in roomair. Your cultures are protected by the cabinet downward airflow, which is filtered by high efficiency particulate air (HEPA) filters. HEPA filters also treat cabinet exhaust air.

30. Working in a Biosafety Cabinet • Use personal protective equipment • Wear a lab coat and gloves. • Plan your experiment in advance • Place everything you need in the cabinet or on an adjacent cart before starting the experiment. • Designate separate areas on the cabinet work surface for clean materials and dirty materials. • Check that the vacuum bottle contains fresh disinfectant (e.g., a one-in-ten dilution of household bleach) and that there is enough space to collect all waste. • Do not use solvents in the presence of open flame.

31. Working in a Biosafety Cabinet • Use careful, sterile technique! • Work with the sash lowered to the indicated level. • Move slowly and deliberately with planned movements. • Do not pass your hands over sterile preparations or containers. • Discharge pipettes against the flask or tube wall to avoid splashes. • Do not re-introduce pipettes into stock containers – use another pipette. • Place disposable pipettes and dishes into biohazard containers and non-disposables into disinfectant.

32. Working in a Biosafety Cabinet • Wash hands before and after procedure. If gloves become soiled, remove them, wash hands and put on new gloves. • Be careful of sharps! • Clean up at the end of the procedure and disinfect surfaces with alcohol. • Before leaving the room, check everything! Then check again!

33. How to Avoid Contaminating your Cultures, You or your Coworkers • Cleanliness!! • You • Wash your hands before and after any procedure • Avoid touching telephones, door knobs, equipment with gloved or dirty hands • Incubators • Wash and disinfect thoroughly on a regular basis • Clean up spills immediately • Check for contaminated cultures regularly and remove immediately • Biosafety Cabinets • Keep free of clutter, particularly to maintain airflow • Keep surfaces clean and disinfected • Clean under the working surface regularly • The Room • Keep counters, floors and equipment clean and uncluttered • Do not overfill biohazard containers • Keep traffic to a minimum and have an organized schedule of use • Organization and Sterile Technique • Proper technique eliminates the need for routine use of antibiotics – save them for when you really need them.

34. Using Autoclaves • Load the autoclave according to instructions. Do not use without prior instruction. • Use loading racks/trays – never place any item directly on the bottom or floor of the autoclave. • Starting the sterilization cycle. Make sure the door of the autoclave is fully closed and the correct cycle has been selected before starting the sterilization cycle. • Unloading the autoclave. Protect yourself from steam and heat by wearing heat-resistant gloves. First, slightly crack open the door. Wait a full five minutes if the autoclave load contains just glassware, and no less than ten minutes when you are autoclaving liquids. Then remove the load and let the glassware stay on the racks 15 minutes before handling the individual pieces.

35. Autoclaving Liquids • Loosen caps. Before loading containers of liquids into the autoclave, the caps must be loosened to avoid having the bottles shatter. • Guard against spills. Use a tray with a solid bottom and walls to contain bottles and catch any spills. Add a quarter to a half-inch of water so the bottles will heat more evenly. • Unload with extreme caution. Always wear a rubber apron in addition to your standard protective equipment when removing bottles that contain liquids from the autoclave. Be alert for a bottle still bubbling - it could explode easily if touched. Let the load stand in an out-of-the-way place for a full hour before handling.

36. Before Leaving the Lab: Turn off: • Gas • Water • Power supplies • Vacuum lines • Compression lines • Heating apparatus

37. Before Leaving the Lab: • Identify and package waste, dispose properly • Put away experimental apparatus, chemicals, etc. • Lock/out and tag/out defective equipment • Decontaminate work surfaces and equipment

38. Before Leaving the Lab: • Return borrowed equipment, apparatus, etc. • Leave lab coat in the lab • Wash • Close and lock door

39. Before Leaving the Lab: • Clean up your mess • Leave the lab safe for everyone • Perform a final check!!!

40. MODULE 2Workplace Hazardous Materials Information System (WHMIS) http://www.hc-sc.gc.ca/hecs-sesc/whmis/msds.htm

41. “No employer may allow the use, handling or storage of a Controlled Productin a workplace unless the product carries a label and a material safety data sheet which meet the requirements of this Act and the regulations and unless the worker has received the training and information required to carry out the work entrusted to him safely”- Article 62.1, An Act respecting occupational health and safety R.S.Q., S-2.1

42. What are Controlled Products?Learn the universal symbols for each class Class A: Compressed Gas Class B: Flammable and Combustible Material

43. Controlled Products Class C: Oxidizing Material

44. Class D Controlled Products Division 1: Materials Causing Immediate and Serious Toxic Effects Division 2: Materials Causing Other Toxic Effects Division 3: Biohazardous Infectious Materials

45. Controlled Products Class E: Corrosive Material Class F: Dangerously Reactive Material

46. Not Classified as Controlled Products: explosives radioactive materials

47. Consult the McGill Laboratory Safety Manual • Learn how to store each class of chemical and separate incompatible classes of chemicals • Learn the limits on storage of hazardous chemicals in the laboratory (e.g., flammable liquids) http://www.mcgill.ca/ehs/laboratory/labsafety/#3.6.3.1

48. Class A: Compressed GasCharacteristics • Gas inside cylinder is under pressure • The cylinder may explode if heated or damaged • Sudden release of high pressure gas streams may puncture skin and cause fatal embolism

49. Class A: Compressed GasPrecautions • Transport and handle with care • Make sure cylinders are properly secured • Store away from sources of heat or fire • Use proper regulator

50. Class B: Flammable and Combustible MaterialCharacteristics • May burn or explode when exposed to heat, sparks or flames • Flammable: burns readily at room temperature • Combustible: burns when heated