Biological Safety Cabinets: Usage Guidelines
Follow these guidelines for working safely in a biological safety cabinet (BSC).
Improper use of a biological safety cabinet can result in contaminated cultures and expose workers to infectious organisms.
When used correctly, a curtain of room air enters the grill at the front edge of the BSC work surface and acts as a protective barrier. The air mixes with the recirculating air stream and passes through a HEPA filter downward toward the work surface, creating a contamination-free zone.
These guidelines aim to preserve the delicate air barrier that protects both the research product and the researcher.
Hands-on training for proper use of a biosafety cabinet is available to researchers. Contact ehsbio@ucsd.edu to schedule a class.
Use the cabinet as intended
- See Biological Safety Cabinets: Overview for a discussion of different types of biosafety cabinets, purpose, placement, certification, and decontamination requirements.
- Do not use the top of the cabinet for storage; this can damage the HEPA filter.
- Keep only necessary equipment or supplies inside the BSC.
- Larger equipment where biohazardous materials are used and/or stored, such as biosafety cabinets, incubators, refrigerators and freezers, must be visibly labeled with the universal biohazard symbol.
Take precautions before beginning work
- Wear personal protection equipment (PPE):
- Labcoat
- Gloves
- Safety glasses
- Place materials in the cabinet before beginning work if possible.
Start up procedures
- Turn off the ultraviolet sterilizer (if so equipped) as soon as you enter the room.
- Turn on all blowers and cabinet illumination lights.
- Allow 5 minutes of operation to purge the system; check the flow alarm system and visual alarm function (if so equipped).
- Decontaminate readily accessible interior surfaces with a disinfectant appropriate for the agents or suspected agents present.
Standard operation procedures
- Never have the ultraviolet light on when working at the cabinet. It can cause eye damage and skin burns.
- Avoid disrupting airflow:
- Minimize movement (especially rapid movements) into and out of the BSC, or in areas near the BSC.
- Do not block the front grill or rear vents with your arms or other materials.
- Work at least 4 inches from the inside edge of the front vent.
- Bunsen burners, other continuous flame devices, or flammable gases are prohibited in biosafety cabinets.
Safer sterilization methods exist, such as touch-plate microburners equipped with a pilot light, small electric "furnaces," or pre-sterilized loops.- See the BSC Open Flames and Flammable Gas Policy for alternatives to continuous flame devices and disinfecting instruments in a BSC.
- Beware of fire hazards associated with vaporized ethanol and isopropanol disinfectants.
Special precautions for ultraviolet (UV) lamps
EH&S Biosafety division strongly discourages UV lamps in BSCs.
National Institutes of Health (NIH), Centers for Disease Control and Prevention (CDC), National Science Foundation/ANSI, and the American Biological Safety Association agree that ultraviolet (UV) lamps are not recommended or necessary for decontamination in BSC’s.
If a UV lamp is used in your BSC, follow the procedures below.
Required:
- Post a warning sign on the front of the BSC indicating the presence of UV light hazards.
- The sign must say CAUTION: Turn off UV light before working.
Be aware of the hazards. Exposure to UV light can cause:
- Painful eye and skin burns
- Damaging exposure levels exist well after the output of the lamp bulb has dropped below the biocidal level.
- Deterioration of some tubing
- This can be dangerous if you're using a touch-o-matic burner with natural gas tubing in a BSC.
Be aware of the limitations:
- Never rely on UV irradiation alone to disinfect a contaminated work area. UV is:
- Not effective on porous materials that are opaque to the light such as wood or foam
- Ineffective if a microbe is protected by dust, dirt, or organic matter
- Affected by the accumulation of dust and dirt on the bulb surface
- Effective only in direct line of site
- UV does not work in shadowed areas, penetrate into cracks or through the grill work of a BSC
- The spill area under the work surface of a BSC is a favorite hide out for fungal spore and hardy bacteria.
- The UV lamp bulb remains lit long after the germicidal effectiveness is gone.
Take precautions during work:
- Turn off UV lamps while the lab is occupied. The stainless steel interior of the BSC can reflect potentially hazardous illumination out of the opening of the cabinet.
- Never have the UV lamp on while an operator is working in the cabinet.
After work is complete:
- Turn the fan off and close the sash, if possible, when the UV lamp is on.
Maintenance:
Because UV lamp intensity (its destructive power) decreases with time, proper maintenance is critical for decontamination purposes. Perform this regular maintenance:
- Clean UV lamp bulbs every other week by turning off the UV lamp then wiping off the surface of the room temperature lamp bulb with 70% alcohol, or in accordance with the manufacturer's instructions.
- Before replacing bulbs, clear the BSC of equipment and material, and disinfect it with 10% bleach and then clean with 70% ethanol.
- Install the bulb with gloved hands to prevent oil build up.
- Disinfect lamp bulbs before disposal as universal waste.
- Check lamp efficiency monthly with a UV meter or monitoring strip.
Shut down procedures
When you are done:
- Decontaminate and remove all items from the interior work area.
- Decontaminate readily accessible interior surfaces with a disinfectant appropriate for the agents or suspected agents present.
- Turn on ultraviolet sterilizer (if so equipped). Note: Never rely on UV irradiation of the work area alone to disinfect a contaminated work area.
- Allow 5 minutes of operation to purge the system.
- Turn off the blower.
Resources
- Decontamination and Sterilization, Office of Research Safety, Dept. of Occupational Health & Safety
- Laboratory Safety: Principles and Practices, Diane O. Fleming, Debra Hunt, 3rd ed., ASM Press
- Appendix A – Primary Containment for Biohazards: Selection, Installation and Use of Biological Safety Cabinets (PDF), CDC (see p. 290)