**Biosafety cabinet (BSC)** are designed to protect the operator, the laboratory environment and work materials from exposure to infectious aerosols and splashes that may be generated when manipulating substances containing infectious agents, such as viruses, bacteria and primary tissue cultures. Unless a chemical hood, a BSC doesn’t protect the operator from volatile toxic chemicals. At LCSB, the BSC are identified with a sticker:
**Biosafety cabinet (BSC)** are designed to protect the operator, the laboratory environment and work materials from exposure to infectious aerosols and splashes that may be generated when manipulating substances containing infectious agents, such as viruses, bacteria and primary tissue cultures. Unlike a chemical hood, a BSC doesn’t protect the operator from volatile toxic chemicals. At LCSB, the BSC are identified with a sticker:
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Basically, the inward air flow from the room forms an air curtain and minimizes the escape of aerosols from the BSC which provides operator protection. The filtered downward air flow protects the samples and the work inside the BSC. About 70% of the air is internally recirculated in the laminar-flow pattern via the main HEPA filter and about 30% of the air is exhausted via a second HEPA filter.
Basically, the inward air flow from the room forms an air curtain and minimizes the escape of aerosols from the BSC which provides operator protection. The filtered downward air flow protects the samples and the work inside the BSC. About 70% of the air is internally recirculated in the laminar-flow pattern via the main HEPA filter and about 30% of the air recirculated into the room via a second HEPA filter.
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The World Health Organization (WHO) published short videos on specifications of BSC:
The World Health Organization (WHO) published short videos on how to properly use a BSC, take a few min to watch them, they are quite short but full of interesting informations:
1.[Introduction to Biosafety Cabinets](https://www.youtube.com/watch?v=KHCT9OJqxPo&t=1)
2.[Preparation Steps Prior to Working in a Biosafety Cabinet (BSC)](https://www.youtube.com/watch?v=4DoHJS8JL4U)
3.[Best Practices for Safe Biosafety Cabinet (BSC) Usage](https://www.youtube.com/watch?v=18QEJUA9XBs)
Please read the following sections to ensure optimal utilization of a BSC:
-[Good practices in a BSC](#good-practices-in-a-bsc)
-[How to deal with a spill in a BSC](#how-to-deal-with-a-spill-in-a-bsc)
-[How to use KOJAIR BSC](#how-to-use-kojair-bsc)
-[How to use LAMSYSTEMS BSC](#how-to-use-lamsystems-bsc)
-[How to use BIOAIR BSC](#how-to-use-bioair-bsc)
-[Maintenance of BSC](#maintenance-of-bsc)
-[How to report an issue with a BSC](#how-to-report-an-issue-with-a-bsc)
Carefully read the below section, it contains specific information for LCSB equipment and practices: how to operate our BSC, how to deal with a biological spill, how to use specific BSC (KOJAIR, LAMSYSTEMS, BIOAIR), how to maintain a BSC and how to report an issue with a BSC.
## Good practices in a BSC
- The user must wear **lab coat** and **gloves** while using a BSC (see [Lab coats](https://howto.lcsb.uni.lu/external/lab/lab-coats/))
- The BSC should be empty when you start working. If material is stored inside the BSC (for instance micropipettes), they must be disinfected when you switch on the BSC
- Be aware that UV-lights act only on surfaces exposed to them, the surfaces in the shadow are not disinfected and a chemical disinfection is still necessary after a UV decontamination
- Be aware that UV-lights act only on surfaces exposed to them, the surfaces in the shadow are not disinfected (ex: below a box, behind a pipette,...). Due to their strong limitation, UV do not replace a chemical disinfection that is still necessary when UV are used. Because of this, we do not recommend to use UV lights as they have strong limitations and give a false sense of safety
-**The BSC must be decontaminated before and after each use**. The interior surfaces should be wiped with an appropriate disinfectant that would kill any microorganisms that could be found in the cabinet. 70% ethanol is not effective against yeast and fungi, you should use Bacillol 30 foam or tissues:
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Corrosive chemicals such as bleach should be avoided, but if used, should be followed with a wipe down of sterile water or 70% ethanol. Wipe the work surface, the inside walls, below the work surface and the collecting pan. Do not use non-sterile paper towels
Corrosive chemicals such as bleach should be avoided (bleach corrodes stainless steel), but if used, should be followed with a wipe down of sterile water or 70% ethanol. Wipe the work surface, the inside walls, below the work surface and the collecting pan. Do not use non-sterile paper towels
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-Allow the cabinet to run for at least 10 minutes after switching on
-Before starting to work, or switching the BSC OFF, allow the cabinet to run for at least 15 minutes. It allows several air filtration to be performed
- Your hands and all the materials that enter in the BSC must be decontaminated with 70% ethanol or Bacillol 30 tissues
- Put Incidin PLUS in the aspiration pump and disinfect the pump tubing:
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- Recycling baskets are found on each floor to discard plastics, empty tip boxes and empty plastic medium bottles **without antibiotics** (if presence of antibiotics, rinse 1X and autoclave the rinsing liquid). However, filtered bottle need to be autoclaved
- Recycling baskets are found on each floor to discard plastics, empty tip boxes and empty plastic medium bottles **without antibiotics** (if presence of antibiotics, rinse 1X and autoclave the rinsing liquid). Filter bottles have to be autoclaved and therefore should not be disposed as recyclable material
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- Resume or terminate work session
- Notify the incident to your laboratory safety coordinator
### **Large spill in a BSC (more than few milliliters of more complicated spill situations)**
The main challenge in this case is also to keep benefiting from the protection offered by the BSC while avoiding to export any contamination outside the cabinet. In case the spill would not be contained in the BSC, priority should be given to treating the part of the spill that lays outside the cabinet while keeping the BSC running.
### **Large spill in a BSC (more than few milliliters or more complicated spill situations)**
The main challenge in this case is also to keep benefiting from the protection offered by the BSC while avoiding to export any contamination outside the cabinet. In case the spill would not be contained in the BSC, priority should be given to treat the part of the spill that lays outside the cabinet while keeping the BSC running.
- Keep the BSC running and keep applying the rules for safe work in a BSC
- If your gloves have been exposed to contamination, change gloves
- Close all open containers
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- Work as clean as possible, please read the [Good practices in a BSC](#good-practices-in-a-bsc) before starting your experiment
- When work is done, disinfect the inner surfaces of the BSC Bacillol 30 foam or tissues. Do not use bleach: it might corrode the stainless-steel plates
- Switch off the light by pressing on <imgsrc="img/img14.png"width="30">
- Allow the cabinet to run for at least 5 minutes with no activity to purge air contaminants from the cabinet
- Allow the cabinet to run for at least 15 minutes with no activity to purge air contaminants from the cabinet
- Stop the ventilation by pressing on <imgsrc="img/img13.png"width="30">
- Close the sash by pressing on <imgsrc="img/img15.png"width="30">
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- Work as clean as possible, please read the [Good practices in a BSC](#good-practices-in-a-bsc) before starting your experiment
- When work is done, disinfect the inner surfaces of the BSC Bacillol 30 foam or tissues. Do not use bleach: it might corrode the stainless-steel plates
- Turn the illumination off
- Allow the cabinet to run for at least 5 minutes with no activity to purge air contaminants from the cabinet
- Allow the cabinet to run for at least 15 minutes with no activity to purge air contaminants from the cabinet
- Stop the ventilation by pressing on “FAN OFF”
- Close the sash using the arrow
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## Maintenance of BSC
**Before and after each use**, the BSC must be disinfected by the user with an appropriate disinfectant that would kill any microorganisms that could be found in the cabinet. 70% ethanol is not effective against yeast and fungi, you should use Bacillol 30 foam or tissues. Corrosive chemicals such as bleach should be avoided, but if used, should be followed with a wipe down of sterile water or 70% ethanol. Wipe the work surface, the inside walls, below the work surface and the collecting pan. Do not use non-sterile paper towels.
**Before and after each use**, the BSC must be disinfected by the user with an appropriate disinfectant that would kill any microorganisms that could be found in the cabinet. 70% ethanol is not effective against yeast and fungi, you should use Bacillol 30 foam or tissues. Corrosive chemicals such as bleach should be avoided (bleach corrodes stainless steel), but if used, should be followed with a wipe down of sterile water or 70% ethanol. Wipe the work surface, the inside walls, below the work surface and the collecting pan. Do not use non-sterile paper towels.
**Once a month**, each BSC should be deeply cleaned with an appropriate disinfectant such as Bacillol 30 and 70% ethanol combined. During this deep cleaning, remove the removable inner surfaces, disinfect the surface under the plates, the inside walls, the grills and the collecting pan. Disinfect and put back the removable inner surfaces. Don’t use corrosive chemicals on stainless-steel plates. Fill the form beside the BSC with the date, your initials and the maintenance performed.
**Once a year**, each BSC is calibrated by the company itself. During the calibration, they check the efficiency of the airflow to ensure that the inflow and the downflow velocities that are displayed by the cabinet are actually measured. They also check the integrity of the HEPA filters. A sticker is placed on the BSC with the due validation date. Before using a BSC, please verify that the BSC has been validated during the past 12 months.
**Once a year**, an operational qualification is realised on each BSC. This qualification consist of :
- a check of the efficiency of the airflow to ensure that the inflow and the downflow velocities that are displayed by the cabinet are correctly measured.
- a check of the integrity of the HEPA filters.
- a measure of the cleanliness of the air via a particle counting
After a successful qualification, a sticker is placed on the BSC with the due requalification date. Before using a BSC, please verify that the BSC has been qualified during the past 12 months.
