Biosafety Cabinet Management

Beyond The Basics

 Table of Contents
Part I - General Guidance
Creating a culture of safety with Sean Kaufman 2
BSCs & fume hoods: What's the difference 8
Differences between American and European BSCs 12

Part II - Class II BSC Type Selection

BSC selection guide 14
To B2 or not to B2? That is no longer the question 18
CETA guide on Type B2 BSCs 22
Type C1, the right compounding BSC 24
Type C1 case study 27

Part III - Use and Accessorizing

4 reasons not to use open flames in BSCs 32
Best practices if a flame is needed 34
UV light use: The good, the bad and the ugly 36
9 things to know before installing an aspiration system 38
 When it counts, count on us.
Labconco has been protecting people since 1925.
We made our first laminar flow biohazard cabinet,
the predecessor to the modern biosafety cabinet,
in 1976. We've accumulated a lot of expertise and
knowledge over the years. This book is a compilation
of articles we think you'll find helpful if your work
involves biological safety.

1 | Introduction labconco.com
 Creating a culture of safety with Sean Kaufman

Safety in the workplace is important regardless where that require Biosafety Level (BSL) 1-3 containment.
you clock in every day. It’s especially important to That just scratches the surface, so we decided to dig
master when your work environment regularly contains a little deeper. We got the input of an outside industry
hazardous particulates, harmful chemical vapors expert to talk about how to create a culture of safety
or radionuclides. in labs, and how that relates to leadership.

Even basic microbiological work has some associated That expert is Sean Kaufman, CEO of Behavioral-
risks. As oftentimes, it is not only necessary to protect Based Improvement Solutions. Sean’s approach
the person sitting at the face of the cabinet but the to the biosafety industry is truly safety-centric, and
science taking place inside on the work surface. It’s with his knowledge has been called upon in high profile
this eye toward protection that labs have rightfully circumstances, including the 2001 Anthrax attacks
turned to using biosafety cabinets (BSCs). This allows and SARS outbreak.
their technicians to properly and safely handle agents

2 labconco.com
 He has testified before Congress about infectious Q & A with Sean Kaufman
diseases. In 2014 he prepared the clinical staff at
Emory Healthcare to respond to the first cases of Q: To start, can you describe a little bit of what you do?
Ebola on American soil. He then traveled to Nigeria A: I’m a behavioral psychologist focusing on infectious
and Liberia to prepare healthcare workers there to disease. Biosafety cabinets are very vital pieces of
do the same. equipment but I focus on human behavior with and
around them. You can spend thousands and thousands
Sean knows his stuff when it comes to biosafety. While of dollars on a cabinet, but all the control can be
his background is diverse, his message is simple: negated in an instant with inappropriate behavior. I
safety matters. We jumped at the chance to ask don’t think the need for biosafety cabinets is any longer
him a few detailed questions. Read on to learn how debated. One, they work. Two, they’re efficient.
to create a culture of safety and why it’s important. Three, they’re clean.

3 | Creating a culture of safety with Sean Kaufman saferbehaviors.com

 They’re one of the best engineering controls we have Ask yourself a few questions
in biosafety today. There are lots of complexities, but • Are you going to be working with animals?
the workforce needs consistent investment in preparing • What type of connection does your science
them to effectively work in a cabinet—that’s where I’m require—are we talking recirculated A2 or
an advocate. thimble connected A2?

• Maybe you’ll need a fully ducted B?

Q: Let’s start dissecting those complexities. There are
so many considerations when it comes to biosafety
cabinet selection. How do you begin to conduct risk From a big picture perspective, we look to make sure
assessments, and why are they important? the cabinet is certified, it is functioning properly and it
A: That’s a big question. When you look at a biosafety passes a smoke test. If all those are in check and you’ve
cabinet, the first and most basic thing you have to do chosen the appropriate equipment, the biosafety
is determine what you’re going to be using it for. engineering aspect has been done for you. Now you
Then, decide how much workspace you need. must operate safely within the enclosure.

4 | Creating a culture of safety with Sean Kaufman saferbehaviors.com

 Q: So, after selecting a biosafety cabinet, what • What type of disinfectant are we using?
comes next? • How are we going to deal with corrosives?
A: It’s absolutely critical to train employees because
• Are we working from clean to dirty?
they are the human interface of the biosafety cabinet,
of that engineering control. Leaders need to train them
how to choose appropriate personal protective The risk assessment list goes on and on, but having
equipment and develop standard operating effective, tested and validated SOPs is critical
procedures (SOPs) that answer real questions. throughout the life of the cabinet. When there
are standard, easy to follow checklists in place
How do we set things up and break them down? In my for routine operations, some of those processes
opinion, by the way, the cabinet should be broken down can be streamlined and the risks minimized.
and cleaned at the end of each night.

5 | Creating a culture of safety with Sean Kaufman saferbehaviors.com

 Q: As a behavioral psychologist, I think you’re more These technicians are dealing with long hours at the
qualified than anyone to answer this next question: bench, and you have to consider posture, arm position
Do you feel ergonomics play a role in safety in the lab? and a number of other behaviors that can affect how
A: Absolutely—but I have my own philosophy on that. safe the work actually is. Adequate preparedness
is critical in the workforce, and that’s a
Leaders do three things. They prepare the leadership responsibility.
workforce. They protect the workforce and they
promote the workforce. When you look at the words Q: Can you explain in greater depth the role of ideal
‘prepare the workforce,’ preparing means that they give leadership in the lab and how that relates to safety?
the workforce the resources they need to do the job A: To me, biosafety cabinets are phenomenal
and to do it well. That’s where good biosafety cabinet engineering devices. However, the minute they come
ergonomics come in. in contact with a human exhibiting human behaviors,

25.6" 24.6"

21" 19"

10" Sash 10" Sash

8" Sash 8" Sash

10" 10"
Worktop 8" Worktop 8"
Height Height
35" 33"

8"

32.5" 30.5"

6" 6"

6 | Creating a culture of safety with Sean Kaufman saferbehaviors.com

 that person better be trained or the cabinet could risk cabinets and contaminating other areas of their labs.
losing its protectiveness. Again, that’s a leadership The workforce is supposed to follow some very clear
thing. You can’t just give someone a biosafety cabinet expectations. Even if they were to follow all of them,
and say, ‘go work.’ You have to train them how to use it. leadership still has a responsibility to the workforce.
Some technicians tend to over-rely on the engineering Guidelines and policies bring us together, but being
aspects of the cabinet. It’s great that they’re working together and doing together are very different things.
in them, but they might botch the glove removal Everyone has to be on-board with safety in order for
process or become lax at disinfecting. That’s how we it to work, and that starts at the top.
have instances of individuals coming out of biosafety

Sean Kaufman spoke before Congress on safety in the lab as it pertains to the anthrax incident of June 2014 at the Center for Disease Control.

7 | Creating a culture of safety with Sean Kaufman saferbehaviors.com

 BSCs & fume hoods: What's the difference

Although they’re both sometimes referred to as “hoods,” If tasked with selecting an enclosure for a lab, below
biosafety cabinets (BSCs) and fume hoods are two are some common points for consideration.
entirely different categories of lab equipment. Both are
designed and built to contain hazards associated with Note that improper selection or use of equipment can
science, but they differ in terms of the precise type(s) yield significant consequences for health, safety and
of protection provided, airflow and suitable applications. operation/function of the lab (and its ventilation
system). Always consult your safety officials for
Fume hoods are ventilated enclosures that remove recommendations based on your specific processes.
chemical fumes and vapors from the lab, providing
personnel protection only. Enclosures for life sciences Protection
utilize HEPA filters and directional airflow to provide Knowing what type of protection you will require is the
environmental, personnel and/or product protection. first step in selecting the proper lab enclosure. BSCs
provide environmental, personnel and product
Both categories can recirculate or exhaust filtered air, protection from hazardous airborne particulates.
depending on your process. While only a subset of Clean benches create a sterile work zone for ‘fragile’
life science enclosures, BSCs, are suitable for work material manipulations. Fume hoods provide only
with hazardous particulates like bacteria, viruses, and personnel protection from hazardous chemical fumes
hazardous drugs. and gases.

FUME HOOD BEST PRACTICES

Proper and undisturbed airflow is key to a safe
and efficient work environment. Do not store
large equipment inside the hood or do anything
that could disrupt the movement of air. Never
put your head inside the hood and keep the
sash closed when not in active use.

Protector ® XStream® Chemical Fume Hood

8 labconco.com/safety-vid
 Fume Hoods: Airflow 2. In a Class II, intake air is drawn safely around
Because fume hoods handle hazardous chemicals (see the operator, sterile air flows downward into the work
applications on page 10), air is drawn past the operator zone and exhaust air is HEPA filtered before it is either
at the front of the equipment. The air passes through recirculated into the lab or released into the atmosphere.
the opening and across the work surface. Finally it
travels through the ductwork before being released to
the outside atmosphere. 3. Class III BSCs are work benches that utilize a
complete physical barrier to contain hazardous work.
Biosafety Cabinets: Airflow Air enters through a single HEPA filter and is exhausted
All three classes of BSCs have one benefit in common: through redundant HEPA filters before being physically
operator safety from hazardous aerosolized/ or chemically treated on its way to the atmosphere.
particulate materials.

1. In a Class I, air is drawn away from the operator,

across the work surface, through a HEPA filter, and
either returned to the lab or vented to the atmosphere.

Purifier ® Logic ®+ Biosafety Cabinet

Class III BSCs can accommodate BSL 4 agents. Class II BSCs can accommodate BSL 4 applications with proper personnel protective equipment.

9 | BSCs & fume hoods: What's the difference bscno-brainer.com

 Fume Hoods: Applications Fume Hoods: Variations
Chemical fume hoods can be used to handle: By-pass chemical fume hoods are the most common
type, operating at a constant air volume. Auxiliary-air,
• Odorous materials reduced air volume and high performance fume hoods

• Toxic gases all fall under the by-pass chemical fume hood umbrella.

• Reactive materials
Of these hoods, high performance hoods operating
• Chemicals that can spatter on a variable air volume (VAV) system are the most
• Carcinogens energy efficient. Hoods exist for special circumstances
as well. Such as those particularly suited to handle
• Flammables
perchloric acid, constructed of acid resistant materials.
• Other toxic and volatile materials Also for radioactive applications, hoods constructed
with extra decontamination features that often require
Biosafety Cabinets: Applications filters in ductwork.
BSCs provide a safe environment for research involving
infectious microorganisms or other hazardous Biosafety Cabinets: Variations
particulates. Depending on the type of cabinet, these Class I BSCs have an open front, operate under
enclosures are suitable for use with agents requiring negative pressure and only provide user protection.
containment in conjunction with BSLs 1, 2 or 3. The most common types of BSCs, Class II, fall under

©Anton Grassl/Esto

Yale University

10 | BSCs & fume hoods: What's the difference labconco.com/safety-vid

 University of Nebraska—Lincoln

three main divisions: Type A, B and C. Class II, Type A for work with or without hazardous chemistry, utilizing
BSCs recirculate air back into the lab. Unless however, installation flexibility to be ducted or not. It relies on
they are canopy connected to the outside as required single pass airflow through a defined area of the work
by the application (work with odorous materials). Class zone and recirculation in outlying areas of the BSC. This
II, Type A1 and A2 BSCs are similar, yet separated by reduces energy demands while providing containment
the minimum average inward air velocity required. Type of biological, particle and chemical hazards.
A BSCs cannot handle work involving radionuclides
or hazardous chemistry. Class III BSCs are completely enclosed and feature
attached rubber gloves.
Class II, Type B BSCs are hard ducted to dedicated
exhaust systems. They are suitable for work with Other types of enclosures. Carefully select the
hazardous chemistry (per a risk assessment), if required proper enclosure for your intended operation. Using
by the microbiological studies being performed in the the appropriate equipment helps protect personnel
interior of the BSC. Type B1 BSCs recirculate a and the lab itself. It can also preserve the integrity of
percentage of air back into the lab while B2 BSCs are your work. Other relevant lab equipment can include
entirely exhausted. glove boxes, clean benches and balance enclosures.
If your application involves nanoparticulates, there are
Class II, Type C BSCs are hybrids of a sort. They're also enclosure options designed to accommodate
reminiscent of Type B1 BSCs. The Type C1 is suitable your specific needs.

11 | BSCs & fume hoods: What's the difference bscno-brainer.com

 Differences between American & European BSCs

National Science Foundation's (NSF) NSF 49 and EN Microbiological challenge testing. NSF 49 allows
12469 are both standards by which biosafety cabinets for testing and acceptance via a microbiological
(BSCs) are manufactured, specifically outlining how challenge. EN 12469 allows for testing and acceptance
they perform. American National Standards Institute via either a microbiological challenge or a KI discus
(ANSI) recognized NSF 49 in 2002, though the standard method test.
has been in use since the 1970s. NSF 49 was also the
basis from which the EN 12469 standard was derived. Downflow velocity. EN 12469 places a requirement on
NSF 49 standardizes and recognizes the different downflow, NSF doesn't. However, NSF requires a larger
subcategories of Class II BSCs (A1, A2, B1, B2, C1), test sample size and requires traceable test equipment
and EN 12469 defines a “general Class II cabinet.” EN for more verifiable testing results. The broad range of
BSCs are nearly equivalent to NSF Class II Type A2 BSC ‘Types’ listed in NSF results in a broader range of
BSCs. The differences between the NSF and EN downflows, as a result of multiple mechanical designs.
standards are explored in this article. Regardless of downflow velocity, both NSF listed and
EN certified BSCs must pass similar protection tests.

Microbiological challenge testing on an NSF listed Class II, Type A2

12 nsf.org
 Inflow velocity. NSF is very specific about inflow Construction. NSF emphasizes cleanability of
requirements. Using a direct inflow measurement (DIM) contaminated surfaces. EN specifies that Class II BSCs
as the primary standard provides repeatable and should be resistant to disinfectants and fumigation.
accurate inflow measurements. EN performs inflow
tests by measuring the airflow above the exhaust Decontamination. EN BSCs frequently include
HEPA filter and back calculating the inflow. NSF A2 gas-tight sash and exhaust cover seals with connection
requires 0.5 m/s, EN requires 0.4 m/s. ports for the administration of decontamination
fumigants such as formaldehyde, vaporous hydrogen
Performance envelope. Defined as the air speed peroxide, or chlorine dioxide by operators. NSF requires
parameters for which the BSC must operate, NSF that BSCs be designed so that they can be
requires that the Class II BSC must function +/-0.025 decontaminated in situ by trained certifiers.
m/s of the inflow and downflow set points. EN does
not have a matching specification, but Class II BSCs Regardless of the differences between the NSF 49 and
must again meet protection testing requirements. EN 12469 standards, the BSC is only as safe as the
individual operating it. Operation, maintenance and
Pressurization testing. NSF requires that every unit certification protocols/schedules should be adhered
manufactured undergo a “soap bubble pressure leak to as prescribed by the BSC manufacturer and the
test” to verify the BSC is of airtight construction. EN user's safety officer.
requires a “type-test” performed by an independent lab
to ensure a design’s leak prevention.

Gas-tight seal in 'open' position on an EN certified Class II, Type A2 Gas-tight seal in 'sealed' position on an EN certified Class II, Type A2

13 | Differences between American & European BSCs EN 12469

 BSC selection guide

There are multiple biosafety cabinet (BSC) classes. But do not offer product protection from contamination,
what are they and how are they different? BSC classes significantly limiting their applications. They use
are categories describing how the BSC works and what unrecirculated airflow away from the operator. Class I
it protects. These “categories” are Class I, Class II and BSCs have a similar airflow pattern to a fume hood but
Class III. they also have a HEPA filter at the exhaust outlet. They
may or may not be ducted outside. Class I BSCs are
Class I. A Class I is defined as a ventilated BSC for safe for use with agents requiring Biosafety Level (BSL)
personnel and environmental protection. Class I BSCs 1, 2 or 3 containment.

Class I

Unfiltered air under

Room air HEPA-filtered air negative pressure

14 bscno-brainer.com
 Class II. A Class II is defined as a ventilated BSC for Type A1. A Class II, Type A1 BSC must maintain a
personnel, product and environmental protection for minimum average inflow velocity of 75 fpm through the
microbiological work or sterile pharmacy compounding. sash opening. They may exhaust HEPA-filtered air back
Class II BSCs are designed with an open front with into the lab, or may be exhaust outside using a canopy
inward airflow (personnel protection), downward HEPA- connection. They are suitable for work using biological
filtered laminar airflow (product protection) and HEPA- agents without volatile toxic chemicals and volatile
filtered exhaust air (environmental protection). These radionuclides, but not for sterile hazardous
BSCs are further differentiated by types based on pharmacy compounding.
construction, airflow and exhaust systems. The types
include A1, A2, B1, B2 and C1. They require all Type A2. A Class II, A2 must maintain a minimum
biologically contaminated ducts and plenums to be average inflow velocity of 100 fpm through the sash
under negative pressure or surrounded by negative opening. Like Type A1 BSCs, they may exhaust HEPA-
pressure ducts and plenums. Type B2 BSCs take this filtered air back into the lab, or may be exhausted
a step further, requiring all biologically contaminated outside using a canopy connection. Type A2 BSCs
ducts and plenums to be under negative pressure or with a canopy connection are safe for work involving
surrounded by directly exhausted negative pressure biological agents treated with minute quantities of
ducts and plenums. Like Class I BSCs, Class II hazardous chemicals. They may also be used with
BSCs are safe for work using agents requiring BSL 1, tracer quantities of radionuclides that won’t interfere
2 or 3 containment. with the work if recirculated in the downflow air.

Class II, Type A1 Class II, Type A2

Unfiltered air under Unfiltered air under

Room air HEPA-filtered air negative pressure positive pressure

15 | BSC selection guide bscno-brainer.com

 Type B1. A Class II, Type B1 must maintain a minimum portion of the BSC (this portion is not marked and
average inflow velocity of 100 fpm through the sash therefore ever-changing as the airflow pattern adjusts
opening. They have HEPA-filtered downflow air with the loading of the BSC’s HEPA filters).
composed mostly of uncontaminated recirculated inflow
air and exhaust most of the contaminated downflow Type B2. A Class II, B2 must maintain a minimum
air through a dedicated duct is directed outside after average inflow velocity of 100 fpm through the sash
passing through a HEPA filter. Similar to Type A2 BSCs, opening. They have HEPA-filtered downflow air drawn
Type B1 BSCs are safe for work involving agents from the lab or the outside air (not recirculated from
treated with minute quantities of toxic chemicals and the BSC exhaust) and exhaust all inflow and downflow
tracer amounts of radionuclides if the chemicals or air to the atmosphere after filtration through a HEPA
radionuclides won’t interfere with the work if recirculated filter without recirculation in the BSC or return to the
in the downflow air. Unlike a Type A2, a Type B1 BSC lab. Because of this, they are sometimes referred to as
is also suitable for work involving minute quantities of 100% exhaust or total exhaust BSCs. Type B2 BSCs
toxic chemicals and tracer amounts of radionuclides are suitable for work involving biological agents
required as an adjunct to microbiology applications as treated with hazardous chemicals and radionuclides
long as the work is done in the directly exhausted rear required as an adjunct to microbiology applications.

Class II, Type B1 Class II, Type B2

Unfiltered air under

Room air HEPA-filtered air negative pressure

16 | BSC selection guide bscno-brainer.com

 Type C1. A Class II, C1 must maintain a minimum Class III. A Class III is defined as a totally enclosed,
average inflow velocity of 105 fpm through the sash ventilated cabinet with leak-tight construction and
opening. Type C1 BSCs are unique in that they can attached rubber gloves for performing operations in the
operate as either a Type A BSC when in recirculating cabinet. Class III BSCs are also called glove boxes. The
mode or a Type B BSC when exhausting. C1 BSCs can cabinet has a transfer chamber that allows for sterilizing
be quickly changed from one mode to the other by materials before they leave the glove box. The cabinet
connecting or disconnecting the exhaust and having is maintained under negative pressure and supply air
the cabinet recertified. The Type C1 also features a is drawn in through HEPA filters. The exhaust air is
work surface with clearly delineated spaces for storage treated with either double HEPA filtration or HEPA
on the sides and work use in the center. The center filtration and incineration. Class III cabinets are safe
work area, when in Type B-mode, has dedicated for work requiring BSL 1, 2, 3 or 4 containment.
direct exhaust so is appropriate for use with
hazardous vapors or radionuclides.

Class II, Type C1 Class III

Unfiltered air under

Room air HEPA-filtered air negative pressure

17 | BSC selection guide bscno-brainer.com

 To B2 or not to B2? That is no longer the question

Regardless of your industry, having the right tools for What if your hammer could do other things you needed
the job matters. That statement goes both ways, too. it to do, though, and actually save you money in the
Of course you want equipment that allows you to long term? The product team at Labconco designed
perform effectively and safely. But you don’t want to a completely new biosafety cabinet (BSC) that can
incur any added expenses for features or protections function in either Type A-mode or B-mode. It’s swiftly
your work doesn’t necessarily warrant. becoming the Swiss Army Knife for lab toolboxes
around the U.S. (like this one at Creighton University).
In other words, if you’re a carpenter, you’re going to
need a hammer—a good, dependable hammer. You Before we explore the Class II, Type C1 BSC, let’s
don’t want to reach for the blunt end of a screwdriver delve into the backstory. It starts by addressing the
in desperation. Or resort to reading an instruction longtime struggle of many labs: determining what kind
manual for a complicated, expensive gadget you’re of BSC—a Class II, Type A2 or B2—is actually required.
too frustrated to use.

18 bscno-brainer.com
 HAZARDOUS CHEMISTRY? It’s important BSC Type Refresher. A2 BSCs recirculate about
to note that although they’re often used 70% of air back into the BSC, while exhausting the
interchangeably in the industry, ‘hazardous remaining 30%. This BSC will protect against
chemicals’ and ‘hazardous chemistry’ do particulates such as bacteria and viruses. Only small
not mean the same thing. quantities of nuisance chemicals may be used in this
BSC when connected to ductwork, since the A2s
For example, if you’re working with a
recirculate (and can concentrate) air within the BSC.
hazardous chemical, you don’t necessarily
Type B2 BSCs exhaust 100% of their air through
need to opt for a Type B2 BSC if that
ductwork. B2s provide the same particulate protection
chemical is diluted enough to be
rendered non-hazardous. as an A2 and can be used with many hazardous
chemicals since all of their air is exhausted, similar to
For many, it’s not always clear. That’s why a fume hood.
a risk assessment must be performed to
determine if the volume and concentration BSC Choice Struggles. The field of lab research is
of the chemistry you’re going to perform is rooted in change and new discoveries. One thing
indeed hazardous. however, remains unchanged: safety matters.

Typical Class II BSC Exhaust System

Zero Pressure Weathercap

Remote Blower

Canopy Connection
(Type A2 only) or
Sealed Connection
(Type B2 only)

Air-Tight Damper

19 | To B2 or not to B2? That is no longer the question bscno-brainer.com

 It matters so much that many customers have the same the remaining 55-70% within the BSC. In contrast,
misconception. They want to buy a Class II, Type B2 airflow through a B2 BSC is similar to that of a fume
because they think they are safer than their Class II, hood, externally exhausting 100% of the air pulled
Type A2 counterparts. through the BSC. The purpose of this design is to
completely remove any toxic chemical vapors or
This is only partly true, and it comes with a hefty radioactive compounds that are generated inside
application caveat. Type B2 BSCs aren’t only safer the BSC.
when your microbiological work uses volatile toxic
chemicals—they are required. B2s bring room air into the BSC through both an
opening in the top of the BSC and through the inlet
In other words, if your application includes hazardous grille. This air flows through an initial HEPA filter and
chemistry, you must have a hard-ducted Type B2 (or then downward through the work area. All of the
C1, which we’ll get to in a moment). contaminated air is then drawn into a negatively
pressured plenum and exhausted through a second
Some risk assessments find that the application in HEPA filter. A dedicated exhaust system and remote
question warrants a Type A2 BSC. A unit that exhausts blower draw all of the filtered exhaust air out of the lab.
about 30-45% of its total airflow while recirculating

Type C1 A-Mode Type C1 B-Mode

Room air HEPA-filtered air Directly exhausted air Recirculated air

Unfiltered air under Unfiltered air under

negative pressure positive pressure

20 | To B2 or not to B2? That is no longer the question bscno-brainer.com

 Regardless of application, Type A2 and B2s can’t be The C1 utilizes a Chem-Zone™ at the center of the
converted into different types of BSCs. An A2 is always dished work surface, which serves as the dedicated
an A2. A B2 is always a B2. In the past, researchers exhaust portion of the BSC when in B-mode. C1 BSCs
who needed both Type A and B functionality would also have two ECM blowers and consistent inflow
have been forced to purchase both types of BSCs. and downflow velocities controlled by Constant Airflow
Profile™ (CAP) technology. The C1 maintains safety
Type C1: Best of Both Worlds. The Class II, C1 BSC for you and your work, even in the event of a
can convert from operation in Type A-mode to B-mode. building exhaust failure.
Running a C1 in B-mode is cost effective, too. It
exhausts much less air than a traditional B2. In fact, C1 BSCs change the way lab biosafety is approached
C1 BSCs exhaust about the same amount of air as an by combining versatility, flexibility and safety. Labs
A2 with canopy connection. C1 BSCs can be tied to no longer have to teeter the line between Type A and
almost any existing exhaust system with sufficient air B or buy multiple units to satisfy multiple needs. The
volume reserve. As such, C1 installation costs are C1 is capable of handling any application. To B2 or not
minimal compared to traditional B2-only uses that to B2 is no longer the question.
require dedicated duct runs and blowers for each BSC.

21 | To B2 or not to B2? That is no longer the question bscno-brainer.com

 CETA guide on Type B2 BSCs

Class II biosafety cabinets (BSCs) use uniform (laminar) What if you need protection from hazards, both
airflow and HEPA filters. They provide protection particulate and vaporous? Total exhaust, Class II, Type
from particulates (biological) hazards for the user, the B2 BSCs may have your attention. Ever progressive
product and the environment. Chemical fume hoods regulations in the pharmaceutical, healthcare and
use exhaust volume and face velocity to maximize research industries have increased the interest and the
containment and removal of hazardous chemicals use of “Class II, Type B2” in lab vocabulary.
and their vapors.

Characteristics of Type B1 and B2 BSCs

Type B1 Type B2
Exhaust System
Required Required
Should have dedicated ductwork Should have dedicated ductwork
Type
and exhaust blower for each BSC and exhaust blower for each BSC
Must pull air through BSC’s exhaust Must pull air through BSC’s exhaust
Function
HEPA filter and then through ductwork HEPA filter and then through ductwork
Volume B1 exhausts approximately 20% less that an A B2 exhausts 100% or more than a Type B1
Negative Static Typically 1.5" H2O minimum Typically 1.5" H2O minimum.
Pressure at BSC Maximum may exceed 4.0" H2O Maximum may exceed 4.0" H2O
Vacuum requirements may increase up to Vacuum requirements may increase up to
Reserve Capacity
2.0" H2O as exhaust HEPA filter loads 2.0" H2O as exhaust HEPA filter loads
Other Characteristics
Must be permanently connected to an Must be permanently connected to an
Cabinet Flexibility
exhaust system to function properly exhaust system to function properly
Cabinet Cost More expensive than Type A More expensive than Type A
More expensive than Type A or B1
Installation Cost Similar to a canopy connected Type A
Higher volumes may require larger ductwork

22 ceta-appguide
 This amplified focus on Type B2 BSCs called for a Among subjects visited are:
central resource to spell out the requirements and
guidelines for B2 BSC installation. These guidelines 1. How to choose the right BSC (Type A vs. B)
maintain maximum safety for the environment, the 2. Theory of operation
product and, most importantly, the users in
3. Characteristics of the various BSC types
increasingly hazardous lab scenarios.
4. Site requirements: cabinet location and exhaust
The Controlled Environment Testing Association’s system parameters, design and construction
(CETA) application guide (CETA CAG-007-2010) 5. Dedicated exhaust requirements for Type B BSCs
addresses this need. It is meant to educate lab
management, support field certifiers and ‘unite’ the
BSC manufacturing industry. The CETA document
illustrates important and often missed information
critical to the correct installation and operation of
Type B BSCs.

Kansas Bureau of Investigation

23 | CETA guide on Type B2 BSCs cetainternational.org

 Type C1, the right compounding BSC

The newest subtype of Class II biosafety cabinet (BSC) or Type B2 BSC is the preferred open-faced
is bucking many trends and decades-old canon. The enclosure. USP <800> maintains that most
Class II, Type C1 provides multiple benefits to the compounding processes require only an A2 unless
modern lab. using chemicals that create hazardous vapors.

Here are four reasons why it best suits the needs of The Class II, Type C1 is safer than a Type B and is
the pharmaceutical compounding facility of tomorrow. operationally more flexible than an A. A C1 BSC can
be ducted in a B-mode. It enhances chemical safety
Convertibility of BSC operation. Requirements for above that of a Type B through use of dedicated
compounding hazardous, sterile drugs (under USP exhaust and superior alarm condition programming.
<797> and <800>) have long held that a total exhaust See Active Protection Protocol.

University of Nebraska—Lincoln

24 labconco.com
 Airflow requirements closer to an A. The large Because a C1 uses airflow channeling, compared to
volume of air needed to exhaust a B2 BSC is a leading mass airflow evacuation found in a B2, it is more
cause of initial lab commissioning failures. Why? suitable for cleanroom installation. Type C1s use
Modular cleanrooms are limited in their overall size, approximately half as much air, making cleanroom air
and B2 BSCs require more than many air handling balance and stability far easier to achieve and
systems and rooms can provide. maintain. Furthermore, since USP <800> requires
containment devices to operate 24 hours a day, the
The following can occur: cost savings of the C1 is even more pronounced over
any B2. Commissioning failures are less likely.
1. B2 installation causes the room to be so negatively
pressurized that doors become difficult to open Bag-in/bag-out exhaust HEPA filter. The airflow

2. There isn’t enough air to feed the B2 so it goes into requirements (and associated cost) of a B2 BSC

an exhaust alarm state typically leads to the selection of an A2 BSC with

exhaust canopy, and understandably so. However,
3. Small rooms with a B2 against one wall become
A2s lack the ability to truly house bag-in/bag-out
proverbial wind tunnels, creating uncomfortable
(BIBO) HEPA filters.
and unreasonable working conditions

25 | Type C1, the right compounding BSC labconco.com

 Standard modular cleanroom is 8' tall (96") from floor
to top of roof panel. An interior height is closer to 95"

Axiom Type C1 Logic+ A2 with Logic+ B2

27.5 + 64.2 = 91.7" Canopy 27.5 + 72.6 = 100.1"
27.5 + 64.2 + 9 = 100.7" 27.5 + 63.3 + 10= 100.8"

9"
10"

64.2" 72.6"
96" 63.3"

27.5" 27.5" 27.5"

Such filter designs are uniquely required for The C1, configured as the B2 just mentioned, has a total
pharmaceutical compounding and research work. height of 7', 8". Other than the penetration required
Like B2 BSCs, the C1 can be factory built with BIBO for the duct stub (needed for any Class II BSC going
exhaust HEPA filter ensuring that the filter can be into a USP <800> cleanroom), no further
changed without contaminating the lab with modification is needed.
hazardous particles.
The Class II, Type C1 BSC addresses a number of
Lower clearance height is an often overlooked significant gaps between A and B BSCs. As
problem for modular cleanrooms. Most standard highlighted above, several of these advantages
cleanrooms have a total height of 8'. Most B2 BSCs significantly benefit the needs and requirements of
on a base stand, with a working height of 30" (seated compounding pharmacies.
height) exceed 8', 4" of total height. As such, specialty
panels are needed in the cleanroom’s ceiling, thus For more information on how the C1 addresses gaps in
increasing project cost. modern design and function, visit BSCno-brainer.com.

26 | Type C1, the right compounding BSC labconco.com

 Type C1 case study

The Challenge: Boost environmental consciousness use for labs without compromising health and safety.”
in research labs without sacrificing safety. So Baxter—who, combined with Chemical and
Sustainability Coordinator, Mary Duda, boast over 23
The innovative culture at Creighton is contagious. years of experience in their field—decided to approach
Faculty and administrators have a history of taking a supervisor with an idea.
initiative when it comes to academics and programs.
The same approach held true when, in 2015, a That supervisor, Associate Vice President of Facilities
committee was created to focus on facility and Project Management Tim P. Norton, championed
improvements that could boost sustainability. the project to fellow members on the Energy Savings
Committee (ESC). Take a look at the use of B2 biosafety
Creighton was not alone, though. John Baxter, Director cabinets (BSCs) in the Beirne Research Tower and Criss
of Environmental Health & Safety (EH&S) at Creighton, Health Sciences Center, as the current functionality
described a “nationwide push by universities and requirements might be met with less energy
research institutions to decrease the energy that they taxing equipment.

Creighton University

27 labconco.com
 Wes Walling is the Director of Mechanical, Electrical The existing B2 BSCs at Creighton were original to
and Plumbing Operations at the university. He's also a the facility. Also, as Walling explained, were used
member of the eight-person ESC. Walling said his team because “in the old days, energy was cheap and they
was looking for projects that would have a five-year just exhausted everything as another way of thinking
or better return on investment (ROI). it was safer.”

For starters, they rolled out LED lighting upgrades and Baxter met with the principal investigators to determine
installed pressure independent control valves for chilled if a replacement in technology would even be possible,
water. One project in particular, though, stood out. based on applications. He learned that none of them
Walling called Baxter’s suggestion to replace the B2 actually needed the full B2 exhaust capability of their
BSCs with more energy efficient and flexible models units at this time. However, they couldn’t rule it out in
“an absolute winner from the start.” the future, either. As a result, running the C1 BSCs
in A-mode would not result in any current changes
The Solution: Replace B2 BSCs with C1s to be used in application. However, if the users did need to work
primarily in recirculating A-mode. Type C1 BSCs with volatile organic compounds (VOCs), they could
can operate in A-mode for use in microbiological convert the units to B2 functionality without
applications. They can also be exhausted for B-mode replacing them.
applications like those that involve hazardous vapors
or radionuclides.

Purifier ® Axiom® C1 BSC

28 | Type C1 case study labconco.com

 The team went even further. They wanted to maximize The result. Saved energy, maintained safety and
the safety of the space in one of the buildings. So, increased functionality. Baxter believes sustainability
Creighton installed a valve system that is monitored is second only to safety when it comes to areas of
and controlled not only by the temperature but also the importance within lab spaces. As such, he said the 11
need for air exchanges in the labs. Thereby they C1 BSCs fit well into Creighton’s facility. After all,
ensured there are no hazardous chemicals in the air. Walling’s projection far exceeds the five-year ROI
The system monitors the lab exhaust air for VOC and benchmark for ESC projects. In fact, his calculations
CO2 particles, and it alerts EH&S if a room exceeds show a project with a 2.6-year ROI and an annual
its parameters. Besides decreasing the amount of energy savings of $52,645.
energy used to condition the room air, this system also
provides an extra level of safety for students and faculty. Functionality and safety have increased as well. “The
design of the bottom pan of the new BSCs provides a
The above is just one example of the emphasis on visual reminder to workers about the three ‘zones’ they
safety at the university. Norton explained the philosophy should be working in,” Baxter said. He was referring to
further, stating, “as the departmental budget manager, the specially designed work space that encourages
efficient use of Creighton University resources is users to practice aseptic technique and work from
critically important, but safety is never compromised. clean to dirty.
This project met both goals and will certainly be a
model for additional enhancements going forward.”

The Axiom ® C1 is less expensive than a

conventional Type B BSC, more adaptable
than an A and can do the work of both

B2

$
A2 or C1
ducted
$
A2 or C1
$ $ not ducted

$ $

$ $ $

29 | Type C1 case study labconco.com

 “The design seems to decrease the amount of clutter “The fact that it automatically goes into sterilization
that is left in the back of the biosafety cabinets. mode for 30 minutes after the sash is closed is terrific,
EH&S is happy about that unintended advantage to too, as it is one less thing we have to worry about
these cabinets.” turning off at the end of the day,” Oldenburg said. “Not
to mention that it extends the life of the UV bulb.”
Peter Oldenburg, Assistant Professor in the Department
of Pharmacology at Creighton’s School of Medicine, From a building perspective, Walling said he couldn’t
said he found the C1 BSCs quieter than the previously be happier with the C1 BSCs Creighton purchased to
utilized B2s. Noting it is a nice feature for an educational contribute to their ongoing energy saving efforts.
setting in particular. Additional features, especially “Maybe it was just the right time for technology to finally
those that increase safety and ease of use, appeal to get in sync with what the lab—and the world—requires
Oldenburg and his lab personnel. for better sustainability.”

The Chem-Zone™ Staging Areas

30 | Type C1 case study labconco.com

 University of Nebraska—Lincoln

31 | Type C1 case study labconco.com

 4 reasons not to use open flames in BSCs

A Class II Biosafety Cabinet (BSC) uses HEPA filtered, 1. The Class II BSC maintains sample protection
laminar (uniform) airflow to provide operator, through delivery of downward laminar airflow over the
environmental and sample protection. For the purpose work area. Hot air rises, so any open flame causes air
of sterility, HEPA filters are typically rated at 99.99+% to rise against the laminar downflow. This creates
efficiency for particles 0.3 micron in size. turbulence and foils the BSC’s ability to protect the
samples in the work area.
Typical microbiological procedures often utilize Bunsen
burners or other open flames to sterilize and/or reduce 2. If an open flame gets too hot, it also has the
cross contamination. However, the use of such open capacity to melt the bonding agent that holds the HEPA
flames inside of a BSC is not recommended for filter media to its frame. This destroys the filter’s
several reasons. effectiveness, leading to loss of containment in the
positively pressured plenum.

3. If the flame goes out and the gas supply valve

remains open, flammable gas would be introduced to
the BSC unabated. In an A2, where up to 70% of the
air within the BSC is recirculated, concentrations of
the flammable gas could reach explosive potential.
This poses a serious risk to not only the BSC, but to
the user and the lab it occupies.

Charred HEPA filter

32 labconco.com
 BSC with fire damage

4. The Centers for Disease Control and Prevention Open flames are not required in the near microbe-free
(CDC) and National Institute of Health (NIH) have also environment of a BSC. On an open bench, flaming the
addressed this in the publication: Biosafety in neck of a culture vessel will create an upward air current
Microbiological and Biomedical Laboratories, 5th that prevents microorganisms from falling into the tube
ed., (BMBL 5th). or flask. An open flame in a BSC, however, creates
turbulence that disrupts the pattern of HEPA-filtered

"Several measures can be taken to reduce the chance air being supplied to the work surface. When deemed

for cross-contamination of materials when working in absolutely necessary, touch-plate micro burners

a BSC. Opened tubes or bottles should not be held in equipped with a pilot light to provide a flame on demand

a vertical position. Investigators working with petri may be used. Internal cabinet air disturbance and heat

dishes and tissue culture plates should hold the lid buildup will be minimized. The burner must be turned

above the open sterile surface to minimize direct off when work is completed. Small electric “furnaces”

impaction of downward air. Bottle or tube caps should are available for decontaminating bacteriological loops

not be placed on the toweling. Items should be and needles and are preferable to an open flame inside

recapped or covered as soon as possible. the BSC. Disposable or recyclable sterile loops
should be used whenever possible."

33 | 4 reasons not to use open flames in BSCs labconco.com

 Best practices if a flame is needed

As outlined by the CDC, if a flame is deemed absolutely Better. While not environmentally friendly, single-
necessary, there are types of equipment widely use plastic loops and lab supplies are friendlier to
available that are safer alternatives to the Bunsen BSC use. If a high capacity autoclave is available,
burner. Some of these use low profile, pedal attenuated consider this a better option to an open flame.
flames while others detect motion.
Best. Electrical sterilizers offer the best solution. They
Good. On-demand, low profile, ‘safety’ burners are can sterilize inoculating loops, needles and culture
available that minimize the impact of a flame on the tubes safely by preventing splashes and aerosols
biosafety cabinet (BSC). Most have control valves that while also eliminating cross contamination. Ceramic
close and pilot lights to burn off extra gas through the tubes dissipate the sterilizing heat, and they do not
valve when not in use. Many have a hands-free option require gas.
that uses motion detection or foot pedal operation.

34 labconco.com
 University of Nebraska—Lincoln

35 | Best practices if a flame is needed labconco.com

 UV light use: The good, the bad and the ugly

For decades, germicidal UV lamps have been used in The Good. UVc is a recommended accessory for your
Class II Biosafety Cabinets (BSCs). The purpose is BSC or laminar flow bench when working with cell
to keep a BSC's interior clean when not in use. Many cultures, PCR or other genetic materials. This is
researchers swear by the use of their UV lamp and because UVc radiation is efficient at breaking up the
would never consider owning or buying an enclosure chemical bonds found in DNA and RNA. Under
without one. Others believe UV lamps provide a false prolonged exposure, these chemical changes lead to
sense of security or don’t see the merit of UV irradiation dysfunctional genetic material and eventual cell death.
(such as the NIH). Germicidal lamps utilize UVc
radiation (typically between 250-290 nm wavelength).
This energy is very good at doing a specific task, but The Bad. Germicidal lamps use low energy radiation—
it is a poor broad-spectrum decontaminant, and here so low that the waves are incapable of penetrating
is why: barriers or of reflecting from most surfaces. This means

36 labconco.com
 that for UV lamps to be effective, the target must be as the only means of “cleaning” their BSC, have a higher
in direct line of sight with the light source. This is bad prevalence of such issues.
if you are using a germicidal lamp as the primary
decontaminant in your lab’s culture enclosure. Two simple rules for using UV lamps

However, this is a good thing for those folks in the 1. If you are using “naked” DNA/RNA or performing
lab with nothing but a sheet of glass between them PCR, UV lamps are excellent at rendering these
and the blue tube of light. That sheet of glass is more materials harmless.
than sufficient to block you from getting an
2. If you are doing anything else with biological
unexpected tan.
material, do not rely on UV lamps alone to keep
The Ugly. There is a strong correlation between using your work area clean. Always wipe down exposed
a UV lamp for primary decontamination and the surfaces with a proper decontaminant prior to
frequent presence of cross contamination. True, cross turning on the UV lamp. Always wipe down
contamination can be caused by a number of factors. exposed surfaces with a proper decontamination
However experience shows that labs utilizing UV lamps after turning off the UV lamp.

37 | UV light use: The good, the bad and the ugly labconco.com
 9 things to know before installing
an aspiration system

Following “Best-Practices,” an aspiration system for a replaced or sterilized. When best practices are ignored,
cell culture lab would look like this: a biological risk is posed to the lab, its central vacuum
system and those working throughout the facility.
A primary flask with retaining ring or support to Here is what you need to know when investigating an
prevent tipping aspiration system for your lab.

B a secondary flask for overflow protection

Protect the user. First and foremost an aspiration
C filter between secondary flask and vacuum
system, improvised or purchased, must be designed
D a vacuum source (CDC) to protect the operators and technicians in the lab.

1. Systems should include efficient biological filters,

However, these guidelines are frequently overlooked
such as 0.2µm, placed between the receiving
in improvised system set-ups. Often times, the
flask or bottle and the pump vacuum system
secondary flask is snubbed. A single flask is placed on
the ground without being anchored, nor placed in an 2. Receiving bottles should be capable of being
overflow bin, and the filter—if included at all—is rarely autoclaved or sterilized

D
C
Culture
A B

CDC. Biosafety in Microbiological and Biomedical Laboratories. Ed. L. Casey MD Chosewood and Deborah E. Wilson. 5th. Washington, D.C.: National Institute of Health, 2009.
Fleming, Diane O and Debra L Hunt, Biological Safety: Principles and Practices, 4th Ed. Washington, DC: American Society for Microbiology, 2006.
VACUUBRAND, Inc. "The New Standard in Aspriation Stations." 2013. Document. 12 September 2013.

38 labconco.com
 Protect the culture. Maintenance of sensitive 7. A primary disinfectant should be used in
cultures is difficult, and cleanliness in the work zone conjunction with the receiving flask/bottle. If using
is paramount in the successful management of bleach as the primary disinfectant in the receiving
culture lines. bottle, ensure that glass is used instead of
plastic containers
3. All surfaces of the system should be smooth and
easy to clean Maintain control. Whether analog, electromechanical
4. Tubing should be cleanable or replaceable or digital, be sure that the level of vacuum you are using

5. If the system will be used by multiple users, then matches your needs.

tubing lines should be isolated all the way to the

receiving flask 8. When working with sensitive cells or loose pellets,
a gentle vacuum should be applied when
6. The receiving flask/bottle should be provided with
removing fluids
an appropriate base for support
9. Large vacuums may be required when removing
larger volumes or for higher flow applications
Protect the lab. Often times, researchers and
technicians have a hyper awareness of their immediate
surroundings. Though important, they must not neglect
the larger lab environment.

BCV Fluid Vacuum Aspiration Systems

39 | 9 things to know before installing an aspiration system labconco.com

 Visit labconco.com/bscebook to learn more.

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