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1. A N U M B A T O O L
L E C T U R E R B I O T E C H N O L O G Y , I U B , I B B B .
Biosafety and Bioethics
(Biological Safety Cabinets)

2. BIOLOGICAL SAFETY CABINETS
Effective means of primary physical containment for biological agents,
especially when aerosols are generated.
 HEPA filters remove particles (min 0.3 microns) with 99.97%
efficiency. (HEPA= High Efficiency Particulate Air)
 There are 3 main classes of cabinets (I, II, III) which provide various
levels of protection. Higher classed cabinets provide more
protection.
1. Level 1 provides personnel and environment protection,
2. Level 2 provides personnel, environment as well as product
protection. There are 4 types of level 2 cabinets, each with
different air circulation configurations.
3. Level 3 cabinets provide the most protection, the user works
through attached gauntlets and there is a pass through, they are
used for BSL level 3 & 4 agents.

3. Biological Safety Cabinets
 HEPA filtered Laminar air flow
 Exhaust
 Personnel, environment & product protection
Laminar flow hoods
 Vertical or horizontal laminar flow
 HEPA filtered air (intake only)
 Product protection only
 Laminar flow clean bench can be used for certain clean activities but
not with cell culture material, drug formulations or manipulating
infectious material. Horizontal or Vertical often used in hospitals
for preparation of IV drugs.
 BSC’s provide protection for various combinations of product,
personnel and environment.

4. Biological safety cabinets VS Leminar flow Hood

5. WORKING SAFELY IN A BSC
Before using the cabinet:
 Ensure BSC is certified
 Turn off UV lamp; turn on fluorescent lamp
 Disinfect work surfaces with appropriate disinfectant
 Place essential items inside cabinet
 Allow the blower to run for 5-10 min before work
 Certification sticker, with date of last certification,
should be located on the front of the BSC. BSCs must
be certified every year.

6. WORKING SAFELY IN A BSC
While using the cabinet
 Ensure material and equipment is placed near the back of the hood,
especially aerosol-generating equipment.
 Do not block any vents
 Use techniques that reduce splatter and aerosols.
 General work flow should be from clean to contaminated areas
 Minimize movement so as not to impede air flow
 Open flame in BSC’s is controversial
 Prepare everything you need ahead of time so you are not moving in
and out of the hood.
 Rapid and excessive movements in the hood affect air flow.
 Open flame is not recommended unless there is absolutely no other
method to perform what you want (i.e. can micro-burners be used).
Contact the ORM before deciding to use open flame.

7. WORKING SAFELY IN A BSC

8. WORKING SAFELY IN A BSC
 After using the cabinet:
Leave blower on at least 5 minutes to purge cabinet
 Remove and decontaminate equipment and
materials
 Disinfect cabinet surfaces
 Turn off blower and fluorescent lamp, turn on UV
lamp15 min.

9. WORKING SAFELY IN A BSC
Maintenance:
 Before and after each use - Work surfaces wiped
down
 Weekly - UV lamp should be wiped clean
 Monthly - All vertical surfaces wiped down
 Annually - UV lamp intensity verified-
 Decontamination with formaldehyde gas-
 Certification* UV light is only effective as long as the
light is well maintained.

10. PERSONAL PROTECTIVE EQUIPMENT
(PPE)
 PPE can become an important line of defence (last
line of defense)
 Responsibility of both the user and the supervisor to
ensure that PPE is worn
 Benefits; possible prevention of exposure, potential
minimization of risk that exposure can occur,
compliments existing controls to enhance personal
protection. PPE protects only the individual wearing
it

11. PPE Criteria for consideration
 Routes of exposure that need to be blocked
 Degree of protection offered
 Ease of use
 Only effective if correctly selected, fitted, used and
cared for, and the individual is trained
 Ensure PPE is removed before leaving the lab
 PPE is often specific for each containment level

12. PPE Footwear Lab Coats/Gowns
 Closed toed shoes should always be worn.
 Booties are worn in some higher containment labs and
animal facilities.
Lab Coats/Gowns
 Long-sleeved, knee length with snaps
 Elastic cuffs
 Back-closing gowns
 Periodic cleaning required
 Closed toed shoes protect against spills and injuries from
dropped sharps.
 Elastic cuffs help prevent spills and contamination

13. PPE
Gloves
 Latex, nitrile & vinyl for work with biological agents.
 Exam gloves should not be reused, change frequently.
 Utility gloves can be disinfected and reused if they show
no sign of degradation.
 Consider tensile characteristics, length of cuff.
 Double gloving.
 BSO can provide assistance finding an alternative for
people with allergies.

14. PPE
Eye & Face Protection
 Goggles, safety glasses to protect the eyes
 Full face shield to protect facial skin.
Respirators
 Only personnel who have been fit-tested and trained
should wear respirators.

15. PRACTICES AND PROCEDURES
 General Safety Guidelines
 Good Microbiological Practice
 Handwashing
 Suspicious Packages
 Specific Procedures
 Centrifuges
 Needles & Syringes and other sharps
 Pipettes
 Blenders, Grinders, Sonicators & Lyophilizers
 Inoculation Loops
 Cryostats

16. GENERAL LABORATORY SAFETY
GUIDELINES
 Mostly common sense, but you must understand the
hazards you face in the laboratory and be adequately
trained to deal with them.
 Basic must be known for all labs. Protect yourself,
others and the environment and indirectly the
product.