Biosafety cabinet
Encyclopedia
A biosafety cabinet — also called biological safety cabinet or microbiological safety cabinet — is an enclosed, ventilated laboratory
workspace for safely working with materials contaminated with (or potentially contaminated with) pathogen
s requiring a defined biosafety level
. Several different types of BSC exist, differentiated by the degree of biocontainment
required. BSCs first became commercially available in 1950.
-filtered as it exits the biosafety cabinet, removing harmful bacteria and virus
es. This is in contrast to a laminar flow
clean bench, which blows unfiltered exhaust air towards the user and is not safe for work with pathogenic agents. Neither are most BSCs safe for use as fume hood
s. Likewise, a fume hood fails to provide the environmental protection that HEPA filtration in a BSC would provide. However, most classes of BSCs have a secondary purpose to maintain the sterility of materials inside (the "product").
Class II cabinets provide both kinds of protection (of the samples and of the environment) since makeup air is also HEPA-filtered. There are four types: Type A1 (formerly A), Type A2 (formerly A/B3), Type B1, and Type B2. Each type's requirements are defined by NSF International
Standard 49, which in 2002 reclassified A/B3 cabinets (classified under the latter type if connected to an exhaust duct) as Type A2. About 95% of all biosafety cabinets installed are Type A2 cabinets.
The principle of operation involves using a fan mounted in the top of the cabinet to draw a curtain of sterile air over the products that are being handled. The air is then drawn underneath the work surface and back up to the top of the cabinet where it passes through the HEPA filters. The air that is exhausted is made up by air being drawn into the front of the cabinet underneath the work surface. The air being drawn in acts as a barrier to potentially contaminated air coming back out to the operator.
The Type A1 cabinet, formerly known as Type A, has a minimum inflow velocity of 75 ft/min. The filtered makeup air is divided equally over the work surface at about two to six inches above the work surface. Exhaust is drawn at the bottom of the cabinet where it rises to the top. At the top of the cabinet, 70% of the air recirculates through the supply HEPA filter, the other 30% of air exhausted through the exhaust HEPA filter. This is due to the relative sizes of the two filters, and dampers typically allow the adjustment of this ratio. This type is not safe for work with hazardous chemicals except when ducted, usually with a "thimble" or canopy hood to avoid disturbing internal air flow..
The Type A2 cabinet, formerly designated A/B3, has a minimum inflow velocity of 100 ft/min. A negative air pressure plenum surrounds all contaminated plenums that are under positive pressure. In other respects, the specifications are identical to those of a Type A1 cabinet.
The Type B1 and B2 cabinets have a minimum inflow velocity of 100 ft/min, and these cabinets must be hard-ducted to an exhaust system rather than exhausted through a thimble connection. In contrast to the type A1 and A2 cabinets, 60% of air from the rear grille is exhausted and only 40% is recirculated. Since exhaust air is drawn from the rear grille, the CDC advises that work with chemicals be conducted in the rear of the cabinet. The Type B2 cabinet is expensive to operate because no air is recirculated within. Therefore, this type is mainly found in such applications as toxicology
laboratories, where the ability to safely use hazardous chemicals is important. Additionally, there is the risk that contaminated air would flow into the laboratory if the exhaust system for a Type B1 or B2 cabinet were to fail. To mitigate this risk, cabinets of these types generally monitor the exhaust flow, shutting off the supply blower and sounding an alarm if the exhaust flow is insufficient.
. Gloves attached to the front prevent direct contact with hazardous materials. These custom-built cabinets often attach into a line, and the lab equipment installed inside is usually custom-built as well.
, and the frequent need to clean and replace the bulb. However, these assertions have been formally disputed in at least one peer-reviewed article which points out that :
When a BSC is serviced or relocated, including replacement of HEPA filters, it must be gas decontaminated. Gas decontamination involves filling the BSC with a poisonous gas, most commonly formaldehyde
gas.
Laboratory
A laboratory is a facility that provides controlled conditions in which scientific research, experiments, and measurement may be performed. The title of laboratory is also used for certain other facilities where the processes or equipment used are similar to those in scientific laboratories...
workspace for safely working with materials contaminated with (or potentially contaminated with) pathogen
Pathogen
A pathogen gignomai "I give birth to") or infectious agent — colloquially, a germ — is a microbe or microorganism such as a virus, bacterium, prion, or fungus that causes disease in its animal or plant host...
s requiring a defined biosafety level
Biosafety level
A biosafety level is the level of the biocontainment precautions required to isolate dangerous biological agents in an enclosed facility. The levels of containment range from the lowest biosafety level 1 to the highest at level 4 . In the United States, the Centers for Disease Control and...
. Several different types of BSC exist, differentiated by the degree of biocontainment
Biocontainment
The concept of biocontainment, also called laboratory biosafety, pertains to microbiology laboratories in which the physical containment of highly pathogenic organisms or agents is required, usually by isolation in environmentally and biologically secure cabinets or rooms, to prevent accidental...
required. BSCs first became commercially available in 1950.
Purposes
The primary purpose of a BSC is to serve as the primary means to protect the laboratory worker and the surrounding environment from pathogens. All exhaust air is HEPAHEPA
High-Efficiency Particulate Air or HEPA is a type of air filter. Filters that are awarded the HEPA accolade are used in various locations, whether in medical facilities, automotive vehicles, airplanes, home filters, or wherever very pure air is sought. The filter must satisfy certain standards of...
-filtered as it exits the biosafety cabinet, removing harmful bacteria and virus
Virus
A virus is a small infectious agent that can replicate only inside the living cells of organisms. Viruses infect all types of organisms, from animals and plants to bacteria and archaea...
es. This is in contrast to a laminar flow
Laminar flow cabinet
A laminar flow cabinet or laminar flow closet or tissue culture hood is a carefully enclosed bench designed to prevent contamination of semiconductor wafers, biological samples, or any particle sensitive device. Air is drawn through a HEPA filter and blown in a very smooth, laminar flow towards...
clean bench, which blows unfiltered exhaust air towards the user and is not safe for work with pathogenic agents. Neither are most BSCs safe for use as fume hood
Fume hood
A fume hood or fume cupboard is a type of local ventilation device that is designed to limit exposure to hazardous or noxious fumes, vapors or dusts. A fume hood is typically a large piece of equipment enclosing five sides of a work area, the bottom of which is most commonly located at a standing...
s. Likewise, a fume hood fails to provide the environmental protection that HEPA filtration in a BSC would provide. However, most classes of BSCs have a secondary purpose to maintain the sterility of materials inside (the "product").
Classes
The U.S. Centers for Disease Control and Prevention (CDC) classifies BSCs into three classes. These classes and the types of BSCs within them are distinguished in two ways: the level of personnel and environmental protection provided and the level of product protection provided.Class I
Class I cabinets provide personnel and environmental protection but no product protection. In fact, the inward flow of air can contribute to contamination of samples. Inward airflow is maintained at a minimum velocity of 75 ft/min. These BSCs are commonly used to enclose specific equipment (e.g. centrifuges) or procedures (e.g. aerating cultures) that potentially generate aerosols. BSCs of this class are either ducted (connected to the building exhaust system) or unducted (recirculating filtered exhaust back into the laboratory).Class II
NSF International
NSF International is an Ann Arbor, Michigan-based not-for-profit public health and environmental organization that provides standards development, product certification, auditing, education and risk management services.-History:...
Standard 49, which in 2002 reclassified A/B3 cabinets (classified under the latter type if connected to an exhaust duct) as Type A2. About 95% of all biosafety cabinets installed are Type A2 cabinets.
The principle of operation involves using a fan mounted in the top of the cabinet to draw a curtain of sterile air over the products that are being handled. The air is then drawn underneath the work surface and back up to the top of the cabinet where it passes through the HEPA filters. The air that is exhausted is made up by air being drawn into the front of the cabinet underneath the work surface. The air being drawn in acts as a barrier to potentially contaminated air coming back out to the operator.
The Type A1 cabinet, formerly known as Type A, has a minimum inflow velocity of 75 ft/min. The filtered makeup air is divided equally over the work surface at about two to six inches above the work surface. Exhaust is drawn at the bottom of the cabinet where it rises to the top. At the top of the cabinet, 70% of the air recirculates through the supply HEPA filter, the other 30% of air exhausted through the exhaust HEPA filter. This is due to the relative sizes of the two filters, and dampers typically allow the adjustment of this ratio. This type is not safe for work with hazardous chemicals except when ducted, usually with a "thimble" or canopy hood to avoid disturbing internal air flow..
The Type A2 cabinet, formerly designated A/B3, has a minimum inflow velocity of 100 ft/min. A negative air pressure plenum surrounds all contaminated plenums that are under positive pressure. In other respects, the specifications are identical to those of a Type A1 cabinet.
The Type B1 and B2 cabinets have a minimum inflow velocity of 100 ft/min, and these cabinets must be hard-ducted to an exhaust system rather than exhausted through a thimble connection. In contrast to the type A1 and A2 cabinets, 60% of air from the rear grille is exhausted and only 40% is recirculated. Since exhaust air is drawn from the rear grille, the CDC advises that work with chemicals be conducted in the rear of the cabinet. The Type B2 cabinet is expensive to operate because no air is recirculated within. Therefore, this type is mainly found in such applications as toxicology
Toxicology
Toxicology is a branch of biology, chemistry, and medicine concerned with the study of the adverse effects of chemicals on living organisms...
laboratories, where the ability to safely use hazardous chemicals is important. Additionally, there is the risk that contaminated air would flow into the laboratory if the exhaust system for a Type B1 or B2 cabinet were to fail. To mitigate this risk, cabinets of these types generally monitor the exhaust flow, shutting off the supply blower and sounding an alarm if the exhaust flow is insufficient.
Class III
The Class III cabinet, generally only installed in maximum containment laboratories, is specifically designed for work with BSL-4 pathogenic agents, providing maximum protection. The enclosure is gas-tight, and all materials enter and leave through a dunk tank or double-door autoclaveAutoclave
An autoclave is an instrument used to sterilize equipment and supplies by subjecting them to high pressure saturated steam at 121 °C for around 15–20 minutes depending on the size of the load and the contents. It was invented by Charles Chamberland in 1879, although a precursor known as the...
. Gloves attached to the front prevent direct contact with hazardous materials. These custom-built cabinets often attach into a line, and the lab equipment installed inside is usually custom-built as well.
Ultraviolet lamps
Neither the CDC nor the U.S. National Institutes of Health recommend the installation of germicidal UV-C lamps in BSCs. The American Biological Safety Association supports this position, citing the safety risk to personnel, shallow penetration, reduced effectiveness in high relative humidityRelative humidity
Relative humidity is a term used to describe the amount of water vapor in a mixture of air and water vapor. It is defined as the partial pressure of water vapor in the air-water mixture, given as a percentage of the saturated vapor pressure under those conditions...
, and the frequent need to clean and replace the bulb. However, these assertions have been formally disputed in at least one peer-reviewed article which points out that :
- There is no cited basis for the need to remove dust and dirt from bulbs
- Properly functioning biosafety cabinets have very clean air so dust is less likely to build up
- Laboratories are generally air-conditioned which eliminates the concern over humidity inhibition of UV effectiveness
- With proper use UV exposure risk to users is very low
- UV disinfection is effective for germicide and virucide as well as inhibiting DNA contamination from PCR
- UV disinfection has the advantage of not leaving residues like physical disinfectants
- The relative safety and risks of UV versus other disinfection techniques (which also entail risks) should be considered
Work practices
As with work on open bench tops, work performed within a BSC must be performed carefully and safely. To avoid contamination and the risk of personnel exposure, the CDC advises investigators to follow best practices to reduce and control splatter and aerosol generation, such as keeping clean materials at least 12 inches (30.5 cm) from aerosol-generating activities and arranging the work flow "from clean to contaminated". In particular, open flames, not necessary within the clean environment of a Class II or III BSC, cause disruption of the airflow inside. Once work inside a BSC has been completed, it is necessary to decontaminate the surfaces of the BSC as with other lab equipment and materials.When a BSC is serviced or relocated, including replacement of HEPA filters, it must be gas decontaminated. Gas decontamination involves filling the BSC with a poisonous gas, most commonly formaldehyde
Formaldehyde
Formaldehyde is an organic compound with the formula CH2O. It is the simplest aldehyde, hence its systematic name methanal.Formaldehyde is a colorless gas with a characteristic pungent odor. It is an important precursor to many other chemical compounds, especially for polymers...
gas.