BIOSAFETY CABINET

MLS 1253 LAB INSTRUMENTATION

LECTURER: SIR SULAIMAN ABDUL HAQ HAZMAN
PRESENTER: VIVIAN KIU WEI CHEE (4222003561)
CONTENT

• INTRODUCTION ABOUT BIOSAFETY

• LEVELS OF BIOSAFETY
• BIOSAFETY CABINET
• TYPES OF BIOSAFETY CABINET
• DO’S & DON’TS WHILE USING BIOSAFETY CABINET
• DECONTAMINATION
DEFINITION OF BIOSAFETY

• Biosafety refers to the principles, practices, and measures implemented to ensure the safe
handling, containment, and control of biological materials to prevent accidental exposure,
release, or unintentional spread of potentially harmful agents. This field encompasses
various activities and procedures aimed at protecting individuals, communities, and the
environment from potential risks associated with the manipulation of biological materials,
including microorganisms, toxins, and genetically modified organisms (GMOs).
KEY COMPONENTS OF BIOSAFETY INCLUDE:

Risk Assessment

Containment Personal Protective Equipment (PPE)

KEY
COMPONENTS
Training and Education OF BIOSAFETY Security

Waste Management Monitoring and Enforcement

L E V E L S O F B IO S A F E T Y
BIOSAFETY LEVEL-1

• Applies to laboratory settings in which personnel work with low-risk microbes that pose
little to no threat of infection in healthy adults.
• Example: nonpathogenic strain of E. coli.
BIOSAFETY LEVEL-1 PRACTICE

• not required to be isolated from surrounding facilities (open table or bench)

• mechanical pipetting only (no mouth pipetting allowed)
• safe sharps handling
• avoidance of splashes or aerosols
• daily decontamination of all work surfaces when work is complete
• hand washing
• prohibition of food, drink and smoking materials in lab setting
• personal protective equipment (PPE), such as; eye protection, gloves and a lab coat or gown
• biohazard signs
BIOSAFETY LEVEL-2

• This applies to laboratories that work with agents associated with human diseases (i.e.
pathogenic or infectious organisms) that pose a moderate health hazard.
• Example: Staphylococcus aureus (staph. infection), HIV
• Personnel working in BSL-2 labs are expected to take even greater care to prevent
injuries such as cuts and other breaches of the skin, as well as ingestion and mucous
membrane exposures.
BIOSAFETY LEVEL-2 PRACTICE

• Appropriate personal protective equipment (PPE) must be worn, including lab coats and
gloves. Eye protection and face shields can also be worn, as needed.
• All procedures that can cause infection from aerosols or splashes are performed within a
biological safety cabinet (BSC).
• An autoclave or an alternative method of decontamination is available for proper disposal.
• The laboratory has self-closing, lockable doors.
• A sink and eyewash station should be readily available.
• Biohazard warning signs
BIOSAFETY LEVEL-3

• This applies to lab works on microbes that are either indigenous or exotic and can cause
serious or potentially lethal diseases through inhalation.
• Examples: yellow fever, West Nile virus, and Mycobacterium tuberculosis.
• Laboratory personnel are under medical surveillance and could receive immunizations for
microbes they work with.
BIOSAFETY LEVEL-3 PRACTICE

• Standard personal protective equipment must be worn, and respirators might be required
• Solid-front wraparound gowns, scrub suits or coveralls are often required
• All work with microbes must be performed within an appropriate BSC
• Access hands-free sink and eyewash is available near the exit
• Sustained directional airflow to draw air into the laboratory from clean areas towards potentially
contaminated areas (Exhaust air cannot be re-circulated)
• A self-closing set of locking doors with access away from general building corridors
• Access to the BSL-3 laboratory is restricted and controlled at all time
BIOSAFETY LEVEL-4

• This applies to lab work with highly dangerous and exotic microbes. Infections caused by
these types of microbes are frequently fatal and come without treatment or vaccines.
• Examples: Ebola and Marburg viruses.
• A BSL-4 laboratory is extremely isolated—often located in a separate building or an
isolated and restricted zone of the building. The laboratory also features a dedicated
supply and exhaust air, as well as vacuum lines and decontamination systems.
BIOSAFETY LEVEL-4 PRACTICE

• Personnel are required to change clothing before entering, shower upon exiting
• Decontamination of all materials before exiting
• Personnel must wear appropriate personal protective equipment from prior BSL levels, as
well as a full body, air-supplied, positive pressure suit
• A Class III biological safety cabinet
BIOSAFETY CABINET

• A biosafety cabinet —also called a biological safety cabinet or microbiological safety

cabinet—is an enclosed, ventilated laboratory workspace for safely working with
materials contaminated with pathogens requiring a defined biosafety level.
COMPARTMENT OF BIOSAFETY CABINET
WORKING PRINCIPLE

• The biosafety cabinet working principle is mainly to suck the air in the biological safety
cabinet outward to keep the biosafety cabinet in a negative pressure state, and to protect
the staff through vertical airflow. The outside air is filtered by an air filter (high-
efficiency particulate air filter, HEPA filter) and then enters the biosafety cabinet to avoid
contamination of the processed samples. The air in the biological safety cabinet also
needs to be filtered by a HEPA filter before being discharged into the atmosphere to
protect the environment.
BIOSAFETY CABINET CLASSES

• Biosafety cabinets are classified into three classes by the U.S. Centers for Disease Control
and Prevention (CDC), each with specific performance characteristics and applications.
• Class I and II Biosafety cabinets are used for Biosafety levels 1 and 2 but, when used
correctly in conjunction with useful microbiological techniques, these provide an
effective containment system for safe manipulation of moderate and high-risk
microorganisms.
• Class III BSCs are most suitable for work with hazardous agents that require Biosafety
Level 3 or 4.
CLASS I BIOSAFETY CABINET

Class 1 biosafety cabinets are primarily used to protect the user and the environment from
exposure to potentially harmful agents, such as microorganisms or chemicals.
They feature a unidirectional airflow that moves from the front of the cabinet towards the
back, creating a barrier between the operator and the materials inside.
These cabinets are designed to provide personnel protection but do not protect the
product or the environment. The air entering the cabinet is typically filtered before release
to prevent contaminants from escaping.
Class 1 cabinets are equipped with an exhaust system that filters the air before releasing it
into the environment. This helps to remove any potential contaminants from the
laboratory.
The cabinet is typically made of stainless steel and features a front opening with a
protective sash. The sash can be raised and lowered to provide access to the work area.
Class 1 cabinets protect the user through inward airflow. The filtered air passes through
the front opening and is then directed away from the user.
Common applications for Class 1 biosafety cabinets include handling biological agents at
Biosafety Level 1 (BSL-1) or BSL-2, as well as various pharmaceutical and clinical
procedures.
CLASS 2 BIOSAFETY CABINET
The Class II biosafety cabinets are classified into three types: Type A1, Type A2, and
Type B2.
Class II cabinets have a laminar airflow that protects the operator and the environment by
preventing the escape of airborne contaminants. The airflow is directed from the top of
the cabinet towards the work surface in a vertical or horizontal pattern, depending on the
specific type of Class II cabinet.
Type A1 and Type A2 cabinets offer operator protection and are suitable for working with
low to moderate-risk agents. They recirculate a portion of the cabinet air and exhaust it
through a HEPA filter.
Type B2 cabinets provide both operator and environmental protection. They have a
dedicated exhaust system that filters all the cabinet air before releasing it into the
environment.
All Class II cabinets have HEPA (High-Efficiency Particulate Air) filters to trap and
remove airborne particles.
Class II biosafety cabinets have a front access opening with a sash that can be raised or
lowered to facilitate the manipulation of materials inside the cabinet.
These cabinets often come equipped with various monitoring systems, including airflow
alarms, filter change indicators, and UV lamps for periodic decontamination.
Class II biosafety cabinets are commonly used for a variety of applications, including cell
culture, microbiological research, pharmaceutical preparation, and other activities
involving hazardous biological materials.
CLASS 3 BIOSAFETY CABINET
 A Class III biosafety cabinet (BSC) is a highly specialized and advanced containment device
designed to provide the highest level of protection for personnel, the environment, and the material
being handled. Class III BSCs are commonly used in laboratories where work involves handling
extremely hazardous agents, such as infectious microorganisms with a high risk of transmission
through inhalation.
 Class III BSCs are enclosed, and the operator works through gloves attached to the front of the
cabinet. The cabinet is designed to provide maximum protection against airborne and liquid
contaminants.
 The Class III BSC is a gas-tight system with a supply and exhaust air system that prevents any
potential release of hazardous materials into the laboratory or surrounding environment.
 Class III cabinets have their own dedicated supply and exhaust air systems, ensuring that the air
inside the cabinet is continuously filtered and kept at negative pressure to prevent the release of
contaminants.
 A Class III BSC typically has a double-door system, where materials and equipment are introduced
into the cabinet through a double-door pass-through box. This minimizes the risk of contaminant
escape.
 Class III BSCs provide the highest level of biocontainment, making them suitable for handling Risk
Group 4 pathogens (agents that pose a high individual risk of life-threatening disease).
 The cabinet is constructed to be gas-tight, ensuring that no air or contaminants can escape the
cabinet during operation.
 The air entering and leaving the Class III BSC is filtered through high-efficiency particulate air
(HEPA) filters to remove particles and microorganisms.
 Class III BSCs often have built-in decontamination systems, such as gas sterilization or vaporized
hydrogen peroxide, to ensure thorough sterilization of the cabinet between uses.
 Advanced Class III BSCs may include integrated monitoring systems for parameters like airflow,
pressure differentials, and filter integrity, ensuring that the cabinet is operating within specified
safety parameters.
HOW TO USE BIOSAFETY CABINET
• Ensure the biosafety cabinet is clean and free from clutter.
• Check that the HEPA filters are functioning properly and have been certified.
• Verify that the airflow indicators and alarms are in working order.
• Place all materials inside the cabinet before turning it on.
• Arrange materials in a way that does not disrupt the airflow pattern.
• Turn on the biosafety cabinet following the manufacturer's instructions.
• Allow the cabinet to run for a few minutes before starting work to establish proper airflow.
• Wipe down all interior surfaces with an appropriate disinfectant before and after each use.
• Keep hands and materials within the cabinet; do not reach over the front opening.
• Work methodically from clean to contaminated areas.
• Minimize movement in and out of the cabinet during use.
• Follow the manufacturer's instructions for shutting down the biosafety cabinet.
• Clean and disinfect surfaces after use.
DECONTAMINATION OF BIOSAFETY CABINET
Decontaminate the BSC before and after every use to prevent cross-contamination. One
method is to use a 1:10 fresh bleach solution followed by a 70% ethanol wipe to avoid
corrosion and achieve good disinfection.
Do not use a spray bottle to apply the disinfectant to surfaces. The solvent vapour
concentrations will be re-circulated in the hood.
Clean or wipe from the clean to the dirty area instead in a circular motion to ensure
complete decontamination.
MAINTENANCE OF BIOSAFETY CABINET
Proper maintenance of a biosafety cabinet (BSC) is crucial to ensure its effective operation and to provide a safe
environment for working with biological materials. Here are some general guidelines for maintaining a biosafety cabinet:
• Follow Manufacturer's Recommendations: Adhere to the specific maintenance guidelines provided by the manufacturer
of the biosafety cabinet. This includes recommended schedules for certification and filter replacement.
• Certification and Testing: Perform regular certification of the biosafety cabinet, which involves testing its performance
and airflow patterns. This is typically done annually by a certified professional.
• HEPA Filter Replacement: HEPA filters trap airborne particles and contaminants. Follow the manufacturer's
recommendations for replacing HEPA filters, and schedule replacements as needed based on usage and certification
results.
• UV Light and Germicidal Lamps: If the biosafety cabinet is equipped with UV lights or germicidal lamps, follow the
manufacturer's guidelines for their replacement. UV lamps should typically be replaced annually, even if they appear to
be working.
• Seal Integrity: Check the integrity of the cabinet seals and gaskets regularly. Replace any damaged or worn seals
promptly to ensure proper containment.
• Airflow Indicators and Alarms: Regularly test and ensure that airflow indicators and alarms are functioning properly.
These indicators provide crucial information about the airflow patterns within the cabinet.
REFERENCE

• https://ehs.umich.edu/wp-content/uploads/2016/08/BSC-WPC.pdf
• https://www.escolifesciences.com/news/optimizing-safety-bsc-installation-and-maintenan
ce-best-practices
• July 15, 2000. Biotechnology. Performance criteria for microbiological safety cabinets.
British Standard EN 12469.
• Marc Dunn, Technical Applications Specialist BSC and Clean Air, Thermo Fisher
Scientific.
THANK YOU FOR
UR ATTENTION