Biosafety Cabinets Standard Operating Procedure

  1. Ready the work area. Move the front window to the maximum opening position and switch the blowers on. Operate cabinet blowers for 15 minutes before beginning work to allow the cabinet to purge or remove particulates from the cabinet.
  2. Wipe down the interior work zone surfaces including the front air grille with 70% isopropyl alcohol.
  3. Wipe down the external surface of items with disinfectant then load the cabinet work zone with all you will need for your work. Pay attention not to overload cabinet’s work zone.
  4. Before you start, make sure you have everything you need. Nothing should be taken out of or put in the biosafety cabinet once you begun working.
  5. Pay attention not to place items onto grille.
  6. During work, make sure your work zone is divided into three areas to create distance between clean area (supplies), working area (working equipment and specimens) and contaminated area (waste containers). Transfer materials from the clean side through the work area to the contaminated side.
  7. Limit the use of sharp instruments, particularly needles and syringes in the biosafety cabinet.
  8. Avoid moving your hands in and out of the biosafety cabinet. When working in the biosafety cabinet use slow and deliberate movements. Avoid side to side arm movements as much as possible.
  9. Enter and exit the cabinet from straight on.
  10. Follow aseptic techniques: keep the open mouth of bottles, tubes and flask parallel to the airflow. Keep the top over petri dishes and tissue culture plates. Transfer viable materials as far back in the cabinet possible and recap vials. Work with one specimen at a time and recap before going on to the next. Discard empty tubes immediately.
  11. After completing your work, run the biosafety cabinet for two to three minutes without activity to purge the air.
  12. Disinfect the external surfaces of all items to be removed from the biosafety cabinet. Spray the items with 70% IPA or ethanol. Leave them in the biosafety cabinet for 10 minutes and then wipe it to clean.
  13. Follow the laboratory guideline for reusable and disposable item decontamination.
  14. Place all wastes in the discard container within the biosafety cabinet.
  15. Remove and discard your gloves and then replace with a new pair. Then remove all the items that have had their surfaces decontaminated.
  16. Remove any waste, disinfect and autoclave or place in the appropriate waste stream.
  17. With the sash window still in the safe position, disinfect all biosafety cabinet surfaces including the work zone and inner side of sliding sash, the work zone side and back walls and the front grille.
  18. Do not spray inside the front grille as only safety trained operators should open and clean under the front grille periodically.
  19. Depending on the laboratory protocol, now you can shut down the cabinet, close the sash window and turn on the UV lamp.
  20. Do not forget to discard your gloves into appropriate bins, wash your hands and remove your coat before leaving the laboratory.

How to Deal…


  1. If a spill occurs while working in the cabinet, do not turn the cabinet off. Remove your outer gloves inside the cabinet as they may have become contaminated. Allow the cabinet to keep running as it will contain aerosols that may have been generated and will purge aerosols through the hepa filter. Check your gown for contamination and discard it if it is dirty. Wash your hand, wear a new gown and gloves.
  2. Completely cover spill with a towel and soap with a 10% bleach solution working from the perimeter to the center of the spill.
  3. After allowing for 15 to 30 minutes for contact time wipe the surrounding exposed surface and equipment with appropriate decontaminant. Contact time will depend on the microorganism and the decontaminant.
  4. Remember to clean the bleach using sterile water afterward as it might get cause discoloration on the stainless-steel surface.


  1. If there are any sharps such as broken cryo ampules pick it up with a pair of forceps and place it in the sharps container. Never pick up sharps with your hand.
  2. After cleaning up a spill, discard your gloves and replace with a new pair.
  3. Allow the cabinet to purge work zone for several minutes.
  4. Make sure all contaminated materials are autoclaved.