safety in the research lab

1. safety in the research lab Merle J Heineke Director, research laboratories Wsu-spokane

2. Resources in Biosafety • Biosafety in Microbiological and Biomedical Laboratories (BMBL) • Issued by the US Department of Health and Human Services: The Centers for Disease Control and Prevention (CDC) and the National Institutes of Health (NIH). This book is the basis for lab biosafety practice and facility inspections. • General lab safety information is not part of BMBL – this information is sourced from other administrative bodies but all is standard practice and regulation in the US. • See The Environmental Health and Safety website. Also the WSU Laboratory Safety Manual and the Laboratory Specific Chemical Hygiene Plan • WSU Flip Chart – each lab should have one posted

3. Definitions • Biohazard: a substance that poses a threat to the health of living organisms, primarily that of humans, and the environment. Biohazards can include medical waste or samples, viruses, bacteria, blood and blood products and toxins. • Biosafety: the application of safety precautions that reduce a researchers risk of exposure to a potentially infectious microbe and limit contamination of the work environment and, ultimately, the community. • Risk Assessment: a process used to identify the hazardous characteristics of a known infection or potentially infectious agent or material, the activities that result in a person’s exposure to an agent, the likelihood that such an exposure will cause a laboratory acquired infection, and the probably consequences of such an infection.

4. Principles of Biosafety • Standard Microbiological Practices and Procedures: personnel should be aware of the hazards they are working with, safe handling techniques, proper lab hygiene, hand washing and proper protective equipment (PPE) • Know what to do in the event of a chemical spill • Hand Washing – soap and running water lathering for 30 seconds • Know the location and use of the eye wash station and shower • N-95 respirator – requires medical clearance and fit testing

5. Laboratory Safety Manual • Laboratory Safety Manual is written for entire campus • No food or drink in the lab • Appropriate clothing (long pants, closed shoes) • Appropriate PPE (lab coat, gloves, safety glasses, respirator) • Wash lab coat regularly • Engineering controls (biosafety cabinets, fume hoods) • Employee accident report

6. Chemical Hygiene Plan • Must be written specifically for each laboratory and be readily accessible to all laboratory staff • Safety Data Sheets (SDS) must be readily available • Standard Operating Procedures (SOPs) must be written for each hazardous agent (hood, PPE, disposal, equipment usage, good lab technique, good housekeeping) • Employee must be trained in the plan, SOP, SDS; training must be documented

7. Waste Collection • Autoclavable Waste – white container, clear bag. Disposable lab ware, gloves, culture plates and media • Biohazardous Waste – red container, red bag. Human body fluids, unfixed tissue • Pathological Waste – orange container, orange bag. Fixed tissue, bodily fluids • Radioactive Waste – yellow container, yellow bag. Radioactive materials • Glassware Waste – labeled plastic containers, bag resistant to broken glass. Place in double lined container. When ¾ full, close bag and transfer to a cardboard box. • Sharps – needles (NOT capped), syringes, blades, suture needles

8. Labels • All reagents must be labeled (contents, lot #, prep date, expiration date). • If small vials are stored within a labeled box, the outer label is sufficient; if removed from the box, the vial must then be labeled. • Hazardous agents must have the appropriate sign indicating the nature of the hazard.

9. Equipment • Your research is only as good as your equipment • Check every day for: Equipment is clean Equipment is at the proper temperature Equipment is certified Is the balance weighing accurately Does the centrifuge actually spin as indicated

10. Equipment – Biosafety Cabinet (BSC) • Primary means of containment when working with infectious or potentially infectious agents. • Class I: do not provide product protection (sterile environment); cannot be used for aseptic technique • Class II: flow prevents aerosols or particulates from escaping the cabinet, captures contaminated air before it reaches the product on the work surface; only types A2, B1, and B2 may be used in manipulation of volatile chemicals in limited quantities • Class III: used for work with highly infectious materials that require maximum protection; there are none of these on the Spokane campus

11. Equipment – Biosafety Cabinet (BSC) • Must be certified on an annual basis • Use optimal sash height to prevent disruption of proper air flow and protection; lifting the sash above the optimal height will cause an alarm to sound • Do not block air intake to the front grille • Keep your movements relatively slow and deliberate • Always wear gloves and a lab coat • Disinfect the work surface before and after usage • The UV light should not work when the sash if up • Return the sash to the down position when done working; turn off the light

12. Equipment – Fume Hood • Designed to work specifically with hazardous and volatile chemicals • Provides no product protection • Use with the sash raised to the optimal height • Clean the work surface before and after each use • Conduct work 6 inches into the work zone • Should not be used for storage of chemicals • Start the blower before beginning work so chemical buildup does not occur • Any equipment placed in the hood should be elevated 2 inches from the work surface • Avoid disruptive air currents • Return the sash to the down position when done working; turn off the light

13. Equipment - -80º Freezers • Check the temperature of the freezer daily • Limit the length of the time the door is open; if open for very long, the freezer will pull a vacuum that prevents you from opening the door until it returns to an acceptable temperature. • Wear the appropriate gloves to prevent frostbite • Care of the freezer: Clean the condenser filters every 2-3 months (main filer and lower filter) Clean the door gasket every 2-3 months; check for punctures or tears Use a plastic scrapper to remove frost buildup; be careful to not puncture or tear the door gasket Clean the condenser at least every 6 months Check the Alarm Battery every 12 months (operates the panel only) Defrost the freezer at least once a year or as needed. • In the event of a problem, contact Merle Heineke (merle.heinke@wsu.edu or x87889)

14. Equipment - Autoclave • Uses a combination of steam, heat and pressure to sterilize • Most significant hazard posed is the risk of steam burns; wear heat resistant gloves when removing materials • Check that the chamber drain is clean and free of debris before each cycle • Use an autoclavable bin to capture all liquids • Position the packages so that they do not touch the sides • After closing the door, check that the screen reads “Jacket Power On” • Select the appropriate cycle (Gravity, Liquid) and the appropriate times • Report any problems to Merle Heineke x87889

15. Equipment - Centrifuge • Centrifugal force separates sample components by density • Keeping the centrifuge clean is critical to prevent cross contamination; wipe with a disinfectant before and after each use • Most important safety procedure is to balance the vials within the rotor; unbalanced rotors by cause extreme vibrations and pose an “explosion” hazard • May also generate aerosols; if working with infectious material, samples must be properly enclosed so that aerosols do not escape. • When finished running, do not open sample tubes outside the confines of a BSC • In the event of a spill or explosion, notify Merle Heineke x87889

16. Equipment – Liquid nitrogen (LN) • Eye protection, gloves and a lab coat should always be worn when working with LN • LN is a serious frostbite hazard • LN can also cause oxygen deficiency as it expands and vaporizes posing the risk of asphyxiation • Cryovials can explode when removed from LN storage due to pressure buildup in the vial

17. Equipment – Compressed gas • All compressed gases are hazardous due to the pressure within the cylinder • All compressed gas cylinders must be tightly secured to prevent them from falling over (attached to wall or floor stand) • Gas cylinders may only be moved on a dolly designed for their transport • Gas cylinders must never be moved while the regulator is in place; only move capped cylinders • Do not attempt to remove/replace the regulator until trained; some gases on this campus post a threat during this process. • If a cylinder needs to be replaced, contact Barry Worden (barry.worden@wsu.edu)

18. Water quality • Domestic water: used for indoor and outdoor household purposes • Deionized water: water that goes through the process of removing ions from it; water is passed through a deionized reaction to remove dissolved particles • Reverse Osmosis water: filters most known water impurities found in domestic water • Purified water: defined by the levels (or lack thereof) of any impurities found in the water. The impurity load of dissolved solids in purified water cannot exceed 10 parts-per-million, and water that meets this threshold is inherently of a higher purity than domestic or filtered water.

19. Contact persons • Merle J Heineke, Director, Research Laboratories ph: 358 7889 or merle.Heineke@wsu.edu Equipment repairs, hood and autoclave maintenance, development of Health and Safety department and programs • Ze Liu, Research Supervisor ph: 358 7633 or ze.liu@wsu.edu Flow Cytometry, Mass Spec, Radiation sampling, dry ice shipments • Megan Chastain, Scientific Assistant megan.chastain@wsu.edu Microscopy, Histology, Imaging • Chad Trent, Industrial Hygienestph: 368 6699 or chad.trent@wsu.edu Chemical waste removal • Barry Worden ph: 358 6842 or barry.worden@wsu.edu Replacement gas cylinders • Pharmacy: Andrew Durganph: Assists Pharmacy labs with general issues

20. Culture of Safety • Safety is everyone’s responsibility. WSU is committed to providing a campus environment that supports the health and safety practices of its community (faculty, students, staff, and visitors) and empowers the community to be responsible for the safety of others. A safe campus environment is a right of employment for all categories of employees. A safe campus learning environment is a right of all involved in education and research. • Good science is safe science. Safety is a critical component of scholarly excellence and responsible conduct of research