To work correctly with the Biosafety Cabinet (BC) it is necessary to follow a series of guidelines before, during and after a certain procedure.
A. Before starting the procedure
– The operation of the BC must be known. The Instruction Manual must be available to the user.
– Activate the UV light for 15 min (ON button on hood, UV bulb button on).
– Turn off the UV light by pressing the UV bulb button and proceed to turn on the BC by pressing the FAN button (fan) and the halogen light (bulb button).
– Check that the extraction system works correctly (never operate without the extraction running).
– Check that the BC flow indicator works correctly and does not present an alarm situation. The value of the flow must be around 0.40 and the exhaust must not exceed 0.50.
– If it is necessary to connect an electronic device such as a bioprinter or scale, activate the plug button.
– Check that there are no open doors or windows, mainly in the environment that could distort the correct functioning of the BC.
– Before use, the work surface must be disinfected by spraying with 70% ethanol or another suitable disinfectant for the agents to be used.
– It is recommended that the material to be introduced into the BC be free of particles, therefore it must be carefully cleaned with 70% ethanol before its introduction into it.
– It is recommended to locate all the material to be used inside the BC before starting work. This minimizes the need to remove the arms during operations and prevents anything from passing into or out of it until the job is finished.
– Before and after having worked in a BC you must wash your hands carefully.
– Long-sleeved gowns with tight-fitting cuffs and nitrile or latex gloves should be used.
B. During the procedure
– The BC should not be used as a warehouse for laboratory materials and equipment.
– It is not advisable to bring into the work area materials that easily emit particles such as: paper, wood, cardboard, pencils, erasers, etc.
-The use of Bunsen burners or similar is not recommended, since their incorrect location inside the BSC can cause deviations and turbulence of the laminar flow and burn the HEPA filters.
– NO object should be placed in front of the front air inlet. The air outlet grill must NOT be blocked.
– It is recommended to work about 15 or 20 cm from the front of the BC. In general, the area of least safety for the user and the product is the 8 cm closest to the front opening.
– Objects should not be placed between the HEPA filter and the area where you are going to work, since shadows and turbulence will be produced. That is, if the containers susceptible to contamination (tubes, culture bottles, etc.) are open, the hand will not pass over them. Nor will they be placed too close to each other to avoid turbulence.
– It is preferable to keep those containers with which you are not working closed to avoid spills and possible contamination. Bottle and tube stoppers are oriented to face up and to the back of the containers to prevent hand rubbing over them.
– In order to preserve the HEPA filters to the maximum, in any type of operation, knocks, the projection of liquids or splashes, perforations, etc., against the protection grid of the same must be avoided.
– Only material that is truly necessary and for immediate use should be brought into the work area. Preferably it will be placed so as to avoid unnecessary movement inside the BC.
– It is recommended to wait 2 to 3 minutes before starting work, when any material has been introduced inside the BC that has laminar flow. This will allow it to reconstitute and purify the possible contamination transported from the outside to the sterile work area.
– It is preferable to use tubes and/or bottles with screw caps instead of cotton caps, since these give off fibers.
– It is advisable to carry out slow movements of the arms and hands inside the BC, since otherwise air currents are created that break the laminarity of the flow and can cause the entry or exit of airborne contaminants.
– The clean utensils must be separated from the contaminated ones, placing them further away from the user and the clean ones closer.
– A container must be placed inside for the collection of sharp objects, to collect used pipettes. No used material should be disposed of outside the BC.
– Spills in the BC must be picked up immediately. You must wait 10 min., from the end of the disinfection, before continuing the work.
– In the event that the BC alarm sounds, all containers containing biological material must be closed immediately, stopping activities until the normal situation returns.
C. At the end of the procedure
– All waste products (culture plates, culture media, samples, etc.) will be removed from the BC in approved, waterproof containers suitable for sterilization, supplied by the hazardous waste management company.
– When the work is finished, all the material must be collected and all interior surfaces must be disinfected with 70% ethyl alcohol or any other suitable disinfectant for the agents used.
– The suction system will be cleaned by absorbing a 10% bleach solution.
– The BC must be kept in operation for a minimum of 5 min. after the job is finished.
– The BC will be closed after use and the UV lamp will be activated for 15 min. The tube of the vacuum system is kept inside the bell.
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