Class II Biosafety Cabinets Guide
Class II Biosafety Cabinets Guide
BIOSAFETY CABINETS
THE BASICS
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NUAIRE CLASS II
BIOSAFETY CABINETS
To run a modern healthcare, biological, chemical, medical research, pharmacy, or pharmaceutical lab, a
manager should understand the capital equipment employed regularly. The more thorough the knowledge of
each device, the more efficiently the lab manager can plan for the implications of deployment, maintenance,
repair, and replacement.
Most equipment in a laboratory performs an active function. fully understood. A 1950 survey reported at the annual meeting
For example, a centrifuge separates liquids, and CO2 incubators of the American Public Health Association noted that 5,000
grow tissue cultures. In comparison, role of a biosafety cabinet, laboratories recorded 1,342 instances of laboratory-acquired
while critical, might seem passive. A biosafety cabinet (BSC) infections. Thirty-nine deaths resulted. Of the 1,342 infections,
does not perform any one direct function, but rather it prevents only 467 had previously been made public. This pattern of
adverse conditions from occurring. BSCs cabinets are meant infection continued in the following decades, as reported by the
to prevent harm to employee health and contamination of the Centers for Disease Control and Prevention (CDC) and National
environment. Some also prevent contamination of the product Animal Disease Center. Such reports may not have addressed
in the work zone. the full scope of harm. Focusing only on human infection may
overlook adverse affects of poor contamination control on the
Biosafety cabinets function by causing air to move in precise
work product of the labs involved.
patterns, as well as filtering air to eliminate particulate matter.
This ability requires sophisticated engineering in such areas as The forerunner of the biosafety cabinet appeared in 1909
the fluid dynamics of airflow, ergonomics of usability, design when a company offered a ventilated hood to prevent infection
of the enclosure, and electrical design of motors, blowers, and with tuberculosis when preparing tuberculin. An enclosed
control systems. cabinet was first mentioned in scientific literature in 1943. By
the early 1950s, the U.S. Army Biological Laboratories at Fort
This e-book focuses on the common types and structures of
Detrick, Maryland developed and implemented sophisticated
BSCs, particularly Class II units, and what lab managers should
containment cabinet technology.2
know about the physics governing BSC functions, construction,
specifications, and practical implications of these properties. Use of containment cabinets began to spread, continuing to the
present. Currently BSCs are considered standard in laboratories
Biosafety Cabinet History to prevent a broad range of potential contaminants.
Development of BSCs was a lengthy process, as the 1991 Clinical
Although BSCs must provide safety for personnel and improved
Microbiology Reviews paper, Biological Safety Cabinetry makes
accuracy in testing and production through controlling airborne
clear.1 The first report of an infection that was the result of
contamination, the terms “safety” and “control of airborne
exposure to materials in a lab came in 1893. The first survey of a
contamination” are relative terms. Is there an acceptable
laboratory-acquired disease, typhoid fever, primarily the result
degree of exposure in a particular instance? Does a standard
of pipette use, began in 1915.
exist by which the effectiveness of BSCs from different
By the middle of the 1900s, the link between laboratory manufactures can be measured?
practices and potential contamination and infection was more
1) Kruse, RH, et. al.; “Biological safety cabinetry”; Clinical Microbiology Reviews; April 1991; pp. 207-241
2) Barbeito, Manuel S. and Kruse, Richard H.; “A History of the American Biological Safety Association Part I: The First 10 Biological Safety Conferences 1955-1965”;
American Biological Safety Association; https://www.absa.org/abohist1.html
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NuAire Class II BioSafety Cabinet Ebook
Class II
Biosafety Cabinet Airflow
Class II
A cabinet with a partially open front for worker access.
The Class II is similar to a Class I in that it uses negative
pressure to keep air from moving through a supply opening and
the access panel. The primary difference between Class I and
Class II cabinets is that a Class II cabinet creates an air barrier
Class I
Biosafety Cabinet Airflow at the front of the cabinet by creating a vacuum using an airfoil
that directs air under the work surface instead of over the work
Class I area. It also filters the air supply to the internal work area and
A cabinet that can be recirculated back into the laboratory or be uses laminar airflow to eliminate turbulence, and the possibility
exhausted through a facility’s HVAC system. It uses negative of cross-contamination within the cabinet. The air splits on the
pressure from an interior blower motor or the external exhaust work surface and is pulled into grills located on the front and
system, creating an air barrier at the front of the cabinet at a back of the cabinet. A portion of air will be drawn from the side
minimum speed of 75 linear feet per minute with the air then of the work area and under the work surface where it is directed
circulating through the cabinet. Negative pressure prevents air up a channel located on the back of the cabinet then either
from spilling back into the lab environment. The cabinet draws re-circulated back into the work zone, exhausted, or a combination
in air from a supply opening typically at the front of the cabinet. of the two depending on a cabinet’s subtype. There are four
Exhausted air from the cabinet is passed through a HEPA filter. subtypes of Class II cabinets. Air is drawn through the cabinet’s
Class I cabinets are designed for low-to-moderate-risk biological front opening at a minimum speed of either 75 linear feet per
substances. They protect personnel and the lab environment minute or 100 linear feet per minute, depending on the subtype.
but not the work product in the cabinet. Some portion or all of the air is either reintroduced into the
cabinet’s work area through a supply HEPA filter or discharged
back into the laboratory or building’s exhaust system through
an exhaust HEPA filter. Class II BSCs are designed for low-to-
moderate-risk biological substances.
3) “Biosafety in Microbiological and Biomedical Laboratories”; Centers for Disease Control; December 2009; https://www.cdc.gov/biosafety/publications/bmbl5/index.htm
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NuAire Class II BioSafety Cabinet Ebook
Class III
of contamination for work located elsewhere in the lab.
Biosafety Cabinet Airflow
To summarize, BSCs are a form of safety control and risk
Class III management that have become standard in virtually every
An enclosed cabinet that protects personnel and environment. lab handling biological materials, whether hospitals and other
Typically designed for biosafety level 4 pathogens. Users place large healthcare providers, testing facilities, pharmaceutical
their hands into mounted gloves that pass through a non-open manufacturers, or other organizations that handle particulate
window and manipulate all work in that fashion. Blowers create matter of low-to-moderate-risk.
a negative pressure of at least 0.5 inches of water gauge. Air
This e-book focuses only on Class II BSCs. Class I cabinets
enters the cabinet through a HEPA filter. Exhaust air passes
have declined in popularity due to the additional benefits
through two separate HEPA filters, or a HEPA filter and an air
Class II versions provide. Class III cabinets are beyond what
incinerator, and finally through ducting to discharge into an
many labs need. From this point forward, if the terms “BSC” or
exhaust system separate from the general laboratory exhaust
“biosafety cabinet” appear without specified type, they refer to
system. Class III BSCs typically do not offer laminar airflow within
Class II cabinets.
the internal work area. Materials pass into the workspace
through a dunk tank on the floor of the cabinet or through a
double-door pass-through box that can be decontaminated
Class II Biosafety Cabinets
BSCs are ingenious in their design. By using relatively simple
between uses. The cabinets are designed for high-risk biological
mechanisms that rely on the physics of fluid dynamics, these
substances.
devices maintain barriers to contamination while still allowing
The Class II BSC has the broadest use and is the most commonly the physical access that allows people to do their work.
found in labs.
The Physics behind BSCs
Need for Class II BSCs BSCs can seem passive because of the most important tool of
BSCs provide protection from airborne contamination by their effectiveness, air, is invisible. To better understand how
particulate matter. The most important need for protection is Class II containment works takes a short review of some basic
the lab technicians operating the BSC. The cabinets prevent physics.
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NuAire Class II BioSafety Cabinet Ebook
each point. When the channel grows larger in cross-section, the How They Work
fluid slows, because volume is directly proportional to both that A BSC is essentially a box with a blower moving air within. A
cross-section and the speed at which fluid moves past the point. transparent panel on the front allows personnel to see inside.
Similarly, when the cross-section shrinks, fluid moves more An opening at the bottom of the panel lets technicians reach in
quickly, so the same volume per time flows past. and perform work. This is known as the window sash.
In the 18th century, Swiss mathematician and physicist Daniel As the blower drives air through the box, the BSC becomes a
Bernoulli applied the conservation of energy to fluid flow that study of fluid dynamics involving the Bernoulli principle. The
led to the Bernoulli Equation and Bernoulli principle: When a moving air column lowers the pressure, creating a negative
fluid flows through a channel, fluid pressure, fluid speed, and pressure area about air pressure in the lab.
channel cross-section are all directly related. So, when the
Even as someone works with hands placed through the bottom
channel size decreases, the fluid speed increases. Increase the
of the window sash, air flows from the lab environment into the
channel size and the fluid speed decreases. Increase the speed
cabinet through that opening and into an air grill at the front of
and leave the channel size the same and the pressure decreases.
the cabinet. The air supply creates a moving air curtain in front
The principle is what lets the airfoil of a plane create lift on the
of the work opening. Because of the negative pressure in the
wing. It also lets a Class II biosafety cabinet operate and protect
cabinet, air does not move outward through the opening into
personnel, product, and the environment.
the lab so long as the person moves hands and arms slowly to
avoid disruption of the air curtain.
Class II Type A2
Biosafety Cabinet Airflow
All or part of the air pulled through the cabinet may be vented
out after passing through a HEPA filter. Depending on the BSC
type and work being done, the filtered air may move into the
lab or can be directed through venting to the building’s exhaust
system, eventually to be carried outside. Although they can
screen particles from the air, the HEPA filters cannot remove
volatile toxic chemical vapors or volatile radionuclides that
might be generated.
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NuAire Class II BioSafety Cabinet Ebook
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NuAire Class II BioSafety Cabinet Ebook
The air supply passes through one HEPA filter creating Type B1
particulate-free air, drawn by a blower. 30 percent of the air Intended for routine microbiological work with generation of
circulated in the cabinet moves through a second HEPA filter minute amounts of chemical vapors, so long as the generation
and is vented out of the cabinet while the remaining 70 percent does not interfere with the flow of air or the work is done at
is directed through the first HEPA filter and recirculates through the rear of the cabinet, where the airflow is 100% exhausted.
the cabinet. Laminar airflow prevents cross-contamination of The cabinet has a minimum inflow speed of 100 linear feet per
items in the workspace. A Class II A1 BSC should not be used minute at the opening on its face. The air supply passes through
with volatile toxic chemicals because the build-up of vapors can a HEPA filter for particulate-free air, drawn by a blower. Of the
create safety hazards. air circulated in the cabinet, approximately 70 percent moves
through an exhaust HEPA filter and is vented out of the cabinet
Type A2
while the remaining 30 percent passes through the supply HEPA
Intended for routine microbiological which either generates no
filter and recirculates through the cabinet. Laminar airflow
chemical vapors. Applications that generate minute amounts
prevents cross-contamination of items in the workspace.
of chemical vapors such as compounding chemotherapy drugs
can be used with an A2 BSC if run with a canopy connection to
an external exhaust. The A2 is similar to the A1 except with
a minimum inflow speed of 100 linear feet per minute at the HEPA Filtered Air
Class II, Type A2
opening on its face, creating greater negative pressure and
Air In-flow 70% Recirculated vs. 30% Exhausted Contaminated Worksurface Air
improving containment.
Contaminated Room Air
ClassHEPA Filtered
II Type A2 Air Class II Type B1
Biosafety Cabinet Airflow
Contaminated Worksurface Air Biosafety Cabinet Airflow
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NuAire Class II BioSafety Cabinet Ebook
Type B2
Intended for routine microbiological work with generation of Again, note that the preceding descriptions are for minimum
minute amounts of chemical vapors. The cabinet has a minimum configurations to meet the requirements of NSF standards.
inflow speed of 100 linear feet per minute at the opening on A cabinet from a particular manufacturer might include more
its face. The air supply passes through a HEPA filter for blowers, larger blowers, higher air speed, or different sizes
particulate-free air, drawn by a blower. Air circulates and of filters.
is drawn through an exhaust HEPA filter to be vented out;
Additionally, a given lab may want capabilities or capacities
100 percent of air is exhausted. Laminar airflow prevents
beyond standard requirements. Because each lab has unique
cross-contamination of items in the workspace.
needs and must determine on how much, if any, air can be
vented to the lab environment versus the volume exhausted
Class II Type B2 from the building, the lab safety officer’s recommendations on
Biosafety Cabinet Airflow
the best type of BSC for the organization are important.
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NuAire Class II BioSafety Cabinet Ebook
The next chapter will discuss the physical construction of a BSC. [B]
Class II Construction
In one sense, it might not seem there should be much to building
a box to fit inside a room and possibly connect to the ceiling
through ducting. However, the simple description belies some
inherent challenges in making a BSC that meets or exceeds
[A]
category specifications.
Durable, high-quality, BSC construction starts with a single piece of type 304
stainless steel. A stainless steel wrap [A] makes up the walls of the cabinet.
Stainless steel sheets [B] are welded in place to make up the top and bottom of
the cabinet. A smaller wrap [C] featuring coved interior corners is welded in place
to make up the back wall of the cabinet. This improves the cabinet integrity by [C]
creating a double sidewall. New work tray supports [D] help minimize the vibration
transfer from the shell of the cabinet to the work zone.
The end result is a monolithic shell of 100% stainless steel without the use of silicone
like many multi-piece cabinets. This provides you with a solid foundation for longer
product life eliminating potential leaks.
[B]
[D]
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NuAire Class II BioSafety Cabinet Ebook
Seam construction is an important detail because over time cabinet in question on a workbench to model the work area and
gaskets and silicon can deteriorate, especially in the presence perform common procedures (though without the introduction
of some substances regularly used in lab work, so riveted joints of material that is hazardous).
may require additional inspection and maintenance.
The work area will help determine the overall size of the
BSC, although models with equal work areas from different
manufacturers can vary significantly in size. For instance, the
front air grill might extend farther into the work zone increasing
the reach of the technician into the safe work area. The design
of the workspace and finish of its interior surfaces are essential
to creating and maintaining laminar flow. Everything within the
workspace should be designed to avoid creating turbulence.
The window sash will have an opening through which users can
place their hands and arms. Working for extended periods of
time with arms held in place can be ergonomically challenging.
Some BSCs have optional platforms that can be put into the
cabinet to give room to rest arms and improve comfort. Plat-
forms are typically placed on the air grill and raised to avoid
interruption of the vacuum.
Venting on the work tray of the work area at the front and back
allow the downward movement of air to maintain laminar flow.
As mentioned earlier, any processes releasing small amounts of
vapor should be performed at the back of the work area to avoid
Caption here?
contamination when the air curtain is disrupted as hands and
arms pass through the front opening.
Work Area and Size
The work area in the BSC is the reason the cabinet, ventilation, Filtration
and control systems exist in the first place. Cabinets vary in size. The entire operation of a BSC revolves around filtration. Below
The larger the work area, the larger the cabinet and air volume. are a series of diagrams for the core ventilation design, by type,
The larger the volume, the more powerful the motor(s) which of Class II BSCs:
are necessary to move the air fast enough to create sufficient
negative pressure. A larger BSC will likely be more expensive, The configurations of the blower(s) and filters can differ, but all
but could increase productivity, aid work flow, and increase the ventilation systems work under the same general principle.
profitability. A motor pushes air through the plenum; a chamber made either
of a flexible material or metal. The plenum forces air through
Work areas are typically between three and six feet (0.9 to 1.8 the supply HEPA filter. The air then moves through the work-
meters) in width, usually with a removable work tray for ease of space in a laminar flow. The airflow splits, passing through grills
cleaning. The size depends on a lab’s needs and the available at the front and back of the work zone and into the bottom
space. It is wise to experiment before deciding on the work area of the cabinet. The air moves up a channel at the back of the
size. Tape or mark off a rectangle approximating the size of the cabinet to the top. At that point the air either is recirculated,
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NuAire Class II BioSafety Cabinet Ebook
going through the same steps above or is exhausted through a The plenum is directly connected to the supply and/or exhaust
HEPA filter either into the lab or through the facility’s exhaust filters and can be either metal or flexible industrial cloth that
system. While all that happens, there is a flow of air through the can act like a balloon and help increase pressure.
air grill at the front of the cabinet. Air is pulled under the work
Another important part of the system is venting. With work
surface to ensure sufficient airflow.
typically performed in an A1 or A2 cabinet, the exhaust from the
The HEPA filters used in a BSC remove particles larger than 0.3 BSC might be safe to release into the lab, having passed through
microns, or micrometers, in diameter, which include bacteria, a HEPA filter. A type B1 or B2 cabinet must be connected to an
viruses, and spores. Any HEPA filter must have an efficiency of at external exhaust system in order to contain vapors, or gases.
least 99.97 percent. A typical life span for a filter is between five
The construction and requirements of BSCs are necessary for
and 15 years, and a new filter will typically cost several hundred
personnel to understand. So are the operation and configura-
dollars. HEPA filters must be replaced in the event of damage,
tion considerations that the next chapter examines.
if an annual certification check finds a leak, or if the filter is
sufficiently dirty that it cannot pass sufficient airflow.
Operation and Configuration
Different manufacturers use different blower configurations. Many aspects of a BSC’s construction are determined by
Some build cabinets with arrays of two or more blowers at a compliance with international specifications, the cabinet type,
given spot rather than a single larger one. This approach the particular size, and safety requirements of the work likely
increases the number of moving parts, increasing the possibility to be performed with the equipment. However, there are more
of a blower failure. Because such a cabinet is designed to considerations beyond the structure.
use the full complement of blowers in an array, failure of one
out of several means the cabinet can no longer offer reliable Control System
containment. Replacement of even one motor requires that the Although the functions of a BSC are simple, for example
BSC be re-calibrated, resulting in lost productivity. blower(s) are on during use, and off when not in use, with
additional running time before and after to purge the
In general, a larger blower motor can move a larger amount chamber — the electrical systems controlling these functions
of air at a higher velocity than a smaller motor. As more can be quite complex.
particulate matter is trapped by a HEPA filter, more pressure
is necessary to move air through the filter. The more power a
motor blower contains the more life one can get out of their
HEPA filters lowering operating costs.
UV Light On/Off
Outlet(s) On/Off
Enter
Lights On/Off
Up Value
Blower On/Off
Down Value
Silence Alarm
with Ring Back Function
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NuAire Class II BioSafety Cabinet Ebook
The main control panel gives an operator the ability to start and Built-in Options
stop the blower(s) and also to monitor two critical functions. Many BSC manufacturers will make custom alterations to
One is the airflow. Technicians and lab managers must know accommodate specific needs for a given lab. That might include
that blowers are active. The protective properties of a BSC any of the following:
require an adequate volume of intake airflow to provide the
necessary negative pressure, maintain the air barrier, and • connections for electrical lines, gas inlets, or equipment
ensure laminar flow. cables
Hence, any BSC needs the ability to monitor the airflow and a • monitoring equipment and associated mounts
way to present the information to the operator. Also necessary
are alarm systems to indicate when something prevents the • computer monitors on the back wall of the workspace
free flow of air, whether blower malfunction, unexpected power
• changes to the work tray and front panel to accommodate
drop, obstructed filter, or constricted chamber or plenum.
a microscope
Just as a variety of BSC models are available from various
• bars and mounts to hang IV bags.
manufacturers, each with its specifics of construction, there
are also different control and monitoring panels on the market.
One model might offer a simplistic display of activity and alerts
via lights. Another might provide more nuanced information
through a digital display. Controls might be analog switches
and knobbed dials or, instead, digital touch panels. There can
also be differences in how the system measures operational
performance. Directly monitoring plenum pressure does not
necessarily offer enough information. A cabinet plenum can be
at the right pressure and still have inadequate airflow to work
properly. Airflow sensors provide a more direct indication of
operational condition.
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NuAire Class II BioSafety Cabinet Ebook
Corridor
LabGard
Ventilation Register
(Blocked Facing LabGards)
Corridor
Casework
LabGard
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NuAire Class II BioSafety Cabinet Ebook
An A1 or A2 cabinet often feeds back into the lab, but either Operation
can be vented out to the building’s exhaust system through Part of the installation and operation of a BSC is deciding what
a canopy. The canopy features gaps where an external fan specific steps should be taken to ensure safety for personnel
located in the exhaust system pulls air from the laboratory and and the environment — and for material in the cabinet if using
captures the BSC’s exhausted airflow. The gaps also allow the a B1 or B2. The safety officer during their risk assessment will
BSC to functional properly if the facility’s exhaust system fails identify specific practices or configurations necessary.
by permitting the exhausted air from the BSC to exit into the
laboratory offering biological and particulate containment only. Decontamination
An externally vented B1 or B2 should be permanently attached Decontamination will be a necessity for any BSC, but the
to the building’s exhaust system through hard ducting. An specific methods will depend on the materials in the cabinet
additional location consideration then becomes whether there and possibly its construction. The manufacturer will have
is clear access to the building exhaust system from where the recommendations of how to safely clean and maintain the
cabinet will stand. equipment.
Your facility must be able to handle the power requirements of The DECON 101 system allows a biosafety cabinet to be isolat-
the BSC. The larger the cabinet the more power is needed to ed from the laboratory using gas-tight seals at the work access
make it function properly (i.e. a 20 amp plug might be needed opening and exhaust port. A supply line allows for the intro-
for larger cabinet widths). The lab manager should work with duction of aerosolized decontaminates, such as formaldehyde
physical plant personnel to examine specs and determine what or H202. An exhaust line allows for the removal of remaining
variations are possible or desirable for the building. aerosols following decontamination.
Class II Type B2
Biosafety Cabinet with permanent exhaust duct Biosafety Cabinet decontamination system
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NuAire Class II BioSafety Cabinet Ebook
Any new BSC will require training for all personnel who might
NuAire Laboratory Equipment Supply
use it. Aspects include set-up, operating the control panel, signs
NuAire manufactures ergonomically designed and engineered
of problems, and procedures to use in case of a problem with
scientific laboratory equipment providing personnel, product
either the cabinet or the building exhaust system.
and/or environmental protection in critical research
The lab manager and safety officer should also adopt a routine environments. NuAire’s extensive line of laboratory equipment
Conclusion
Understanding the structure, configuration, and deployment
of Class II biosafety cabinets is important for labs that need to
install and use them. An introduction like this one can help, but
only so much. Biological Safety Cabinets Animal Research Products
Polypropylene Centrifuges
Fume Hoods & Casework
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