[Video] Chapter 3: Proper Preparation - The 4 Steps to Take Before You Begin Work in Your Biosafety Cabinet

December 14, 2015

Biosafety Cabinet PreparationStep 1: Develop a Standard Operating Procedure and Manual

Each laboratory should develop or adopt a bio-safety operations manual which identifies the hazards that will or may be encountered, and specifies practices and procedures designed to minimize or eliminate risks.

Laboratory personnel must receive appropriate training on potential hazards associated with the work involved, including the necessary precautions to prevent exposures, and how to react to exposure.

The laboratory director should establish policies and procedures so that only those who have been advised of the potential hazard and meet specific entry requirements – such as immunization – are permitted to enter the laboratory or animal rooms.

Step 2: Prepare the Biosafety Cabinet (BSC)

Turning on the Cabinet

There’s more to safely turning on your BSC than merely flipping a switch.  Cabinet blowers should be operated at least three-to-five minutes before beginning work to allow the cabinet to "purge." This purge will remove any particulates in the cabinet.  Make sure the work surface, interior walls, and the surface of the window are disinfected.  Also, the UV  germicidal lamp is a valuable ally, but should not be solely relied on to provide a clean and disinfected work area. This goes for your work materials as well.

Disinfecting Work Materials

The surfaces of all materials and containers placed into the cabinet should be disinfected. This simple step will reduce the introduction of mold spores and thereby minimize contamination of cultures. You can further reduce the presence of microbes on materials used in your Biological Safety Cabinets by periodically decontaminating incubators, refrigerators, and nearby equipment.

Step 3: Prepare Experiment Equipment

It is important to prepare a written checklist of materials necessary for a particular activity. Know what you are working with.

Place everything needed to complete your job inside the cabinet before beginning work. This minimizes the number of arm-movement disruptions across the fragile air barrier of the cabinet. If possible, keep physical activity in the lab to a bare minimum.  Too much movement past the cabinet can disturb the cabinet’s airflow.

Air intake to your BSC is also significant. Remove any items from the intake grills that might block or disrupt the air supply – and be sure the front grill is not blocked, covered or obstructed.

All materials should be placed as far back in the cabinet as possible, toward the rear edge of the work surface and away from the front grill of the cabinet, but still within reach.  Similarly, aerosol-generating equipment should be placed toward the rear of the cabinet to take advantage of the air split.  It is important to wait for at least one minute before working with material inside the cabinet, allowing the cabinet to stabilize and “air sweep” the hands and arms, removing contaminants. 

Make sure extra supplies such as gloves, culture plates or flasks are stored outside the cabinet. Only the materials and equipment required for the immediate work should be placed in the cabinet’s work area.

Handling Pipettes

Upright pipette collection containers should not be used in Biological Safety Cabinets or placed on the floor outside the cabinet. The frequent inward/outward movement needed to put objects in these containers is disruptive to the integrity of the cabinet air barrier and can compromise both personnel and product protection. Only horizontal pipette discard trays containing an appropriate chemical disinfectant should be used within the cabinet.

Mouth pipetting is prohibited; only mechanical pipetting devices must be used.

All procedures must be performed carefully to minimize the creation of splashes or aerosols.

Step 4: Dress Properly

Proper dress is another essential for the protection of personnel, product, and the environment.

Laboratory coats should be worn buttoned over street clothing, protective eyewear should be on at all times and latex, or nitrile gloves are necessary when handling culture, contaminated surfaces, or equipment.

Finally, laboratory equipment and work surfaces should be decontaminated with an appropriate disinfectant on a routine basis; This is important after work with infectious materials and especially after overt spills, splashes, or contamination by infectious substances.

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