Appendix F
Biological Safety Cabinet Operating Procedures:

______________/__________
(Lab Building/Room Number)

Refer to CDC-NIH Biosafety in Microbiological and Biomedical Laboratories, Biosafety Level 2 (BSL 2), C. 1, the WSU Laboratory Safety Manual, and the WSU Safety Policies and Procedures Manual (SPPM) section S80.10.1.

(i.e.: Proper use of biological safety cabinets should be referenced to the cabinet owners/operators manual.)

Proper use of the Biological Safety Cabinet from the Baker Company's Owner Manual.

Start up Procedure

1. If the unit has not been left running continuously, first turn the blower switch to on. Make sure that you have cabinet airflow either by listening for blower sound, feeling the airflow across your fingers, or checking the minihelic gauge for a reading other than zero. The reading on the gauge should be consistent with the last time the unit was on.
2. Turn on the fluorescent light. Never leave the ultraviolet light on while there is anyone in the room.
3. Check to see that the drain valve is in the closed position or the drain coupling is capped.
4. Wipe the interior area of the cabinet with a surface disinfectant. Virex will typically be used since bleach can corrode and pit the steel surface. If bleach must be used, clean the surfaces afterwards with a detergent and rinse with tap water to prevent corrosion.
5. Place all material to be used for the next procedure inside the cabinet. It may be necessary to disinfect the exterior of these materials. Everything required (and nothing more) should be placed in the cabinet before beginning your work so that nothing passes in or out through the air barrier until the procedure is completed. Implements should be arranged in the cabinet's work area in logical order so that dean and dirty material are segregated, preferably on opposite sides of the work area. Note that the more equipment that is in the cabinet, the greater will be the air turbulence it causes. Equipment that rotates, vibrates, or heats should be avoided, or if must be used then these should be placed towards the rear of the cabinet.

   Blocking the front and rear grilles must be avoided. If wipes are used, be sure to keep them away from the grilles.

6. Make sure that the vertical sliding viewscreen is open exactly eight inches.
7. After the cabinet has operated for three minutes with the viewscreen in proper position, you are ready to begin.

Work in the cabinet

1. Hands and arms should be washed before and after work in the cabinet. Gloves should be worn when working with infectious material.
2. Perform all work in the depressed area of the solid work surface.
3. Work with slow movements. Do not move things (arms included) in and out through the air barrier.
4. Open and close doors as little as possible when hood is in use. Such activity causes air disturbances that might interfere with cabinet airflow.
5. Discard pipettes in a suitable container within the cabinet.
6. Flames are usually not necessary, and because they disturb airflow are discouraged. Any use of flames in the hood must have prior approval.
7. Never operate the hood while a warning light or alarm is on.
8. After a procedure has been completed, all equipment which has been in contact with the research agent should be enclosed, and the entire surface decontaminated. Trays of discarded pipettes and glassware should be covered. The cabinet should then be allowed to run for at least three minutes with no activity so that the airborne contaminant will be purged from the work area. All equipment is then removed from the cabinet.
9. Decontaminate the interior surface of the cabinet. In particular check for areas of spilled or splashed media. Never use the cabinet to store supplies or laboratory equipment.
10. If the cabinet is turned off, close the sash completely.
11. If an accident occurs during cabinet operation which causes spills and spatters of an infectious agent around the work area, decontaminate all items and surfaces before any items are removed (the fewer items under the hood the easier it is to clean!).

    If the spill of an infectious agent was enough to create puddles or liquid in the drain pan then the following procedure should be followed:

1. Leave the cabinet running and close the viewscreen for about 5 minutes. This will allow aerosols to settle before starting cleanup.
2. The drain pan should be flooded with 109.0 liters of 10% bleach or virex. Leave the disinfectant in the pan for at least 15 minutes, longer if the spill involved a lotof protein or protein-like material. The disinfectant then needs to be drained out and the surfaces thoroughly cleaned with water to prevent corrosion.

Ultraviolet Light

1. The UV light is typically turned on after all operations involving a biosafety level two infectious agent and left on for 15-30 minutes.
2. Eyes and skin should not be exposed to UV light. It should be turned on only when no one is in the room. It should never be left on over night.
3. UV light cannot be relied upon as the sole decontaminating agent. Surface decontamination is always essential.

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