Quick Protocol: How to Work Safely and Effectively with a Biological Safety Cabinet / Culture Hood

Before using any Biological Safety Cabinet (BSC) for the first time, have a person with working knowledge of the machine give you an overview of how to use the cabinet.

Different labs have different protocols in regards to running the cabinet, disinfecting the cabinet, determining which pathogens that may be used in the cabinet and shutting down the cabinet.  This guide is specifically for class II BSCs.

Starting up a BSC.

1. If required, sign up for the cabinet for duration of use including start up and shut down times.
2. Biosafety cabinets are inspected once a year – make sure annual inspection is up to date.
3. If on, turn off UV light, raise sash to level indicated on the outside of the cabinet, turn power, blower, and lights on and let run for >15 minutes prior to putting anything in the cabinet.
4. Check Magnahelic gauge to make sure air flow is adequate.
5. Disinfect work surfaces inside the cabinet with appropriate disinfectant (70% ethanol is commonly used) by spraying all surfaces and wiping down with a kimwipe.  Be sure to move and wipe under any items stored in the cabinet.
6. Spray off all items to be used in the cabinet with 70% ethanol, wipe with a kimwipe and place in the middle of the work surface area in the cabinet.  To minimize movement into and out of the hood, set up the hood with everything needed for the experiment.

Working in the BSC.

1. If required, put on lab coat.  If lab coats are worn in the hoods, they should be reserved for hood use only.
2. Spray gloves with 70% ethanol and rub hands together to disperse alcohol all over gloves.
3. Adjust lab chair so you can easily reach center and back of hood.
4. Set up layout of hood to provide easy access to all items needed for task.
5. Always work with hands towards center and back of hood.
6. Loosen lids of bottles and tops of pipet boxes to allow easy access.
7. Segregate work into clean and contaminated areas.
8. If using a vacuum line for aspiration, ensure vacuum is adequate and liquids do not back-wash into hood.
9. If using a vacuum line and flask for aspiration, make sure trap does not overflow.
10. If you have to leave the hood while working, close all pipet tip boxes, put covers on bottles and close lids of dishes prior to leaving the hood.
11. Resterilize gloves with ethanol prior to reentry.

Shutting down a BSC.

1. Disinfect all contaminated lab ware.
2. Remove all supplies brought into the hood.
3. Place biohazardous materials in appropriate receptacles.
4. If vacuum flask was used for aspiration, make sure lines and trap are properly disinfected and trap is emptied when full.
5. Disinfect work surfaces with appropriate disinfectant.
6. If using bleach as a disinfectant, wipe surfaces with water or ethanol after bleach solution is removed to prevent pitting of surfaces.
7. Turn off lights, close sash and turn on UV lamp if used.

Any questions? Let me know in the comments!