1. Working Safely In Your Biological Safety Cabinet

2. Training Overview Part I. HEPA & ULPA Filters
Part II. Different Types of Cabinets
Class I, Class II and Class III BSC
Differences between A2 and B2 BSC
International Standards
Part III. Biosafety Cabinet Certification
Part IV. Common user mistakes
Part V. Demonstration on how to use a BSC

3. HEPA & ULPA Filter
HEPA: High Efficiency Particulate Air ULPA: Ultra Low Penetration Air
Important definitions:
- HEPA: 99.99% - at 0.3 microns
- ULPA: % - at 0.12 microns
Note: The “classical” definition of HEPA filter is
99.97% at 0.3 microns, but nowadays all
BSC and LF in US use 99.99% at 0.3 m
European definition 99.95% at MPPS EN 1822
Classifications to speak about H %
H %, U % at MPPS

4. Particle Size Comparison
Air contains various foreign materials
Atmospheric dust (mixture of mist, fumes, dry granular particles, fibres, soot, smoke, quartz, clay, traces of decomposed animals and plants)
Definition of aerosol (floating particles in air)
Particle size measurements
Micrometer = micron = 1 thousandth of a milimeter = 10 power minus six of a meter

5. HEPA/ULPA Capability Removes a broad range of airborne contaminants:
Fine dust
Smoke
Bacteria (typical size: 500 to 0.3 micron)
Soot
Pollen
Radioactive particles
Impurity ion -> can affect Integrated Circuit
speed

6. Types of Cabinets Biohazard Safety Cabinet (BSC)
Laminar Flow Cabinet (LFC)
Fume Hood
During review of classifications explain european first (This is sort of like a review of the introduction), then go on to the more detailed american classification
Mention example of british cabinet double exhaust
Also Italian / Australian cabinet style double fan
Also remember to draw out all the airflow in great detail

7. Laminar Flow Cabinets Product protection (no personnel protection)
Not for biohazard agents or chemical fumes

8. Biosafety Cabinets Class I BSC: Personnel and Environment Protection
Class II & III BSC: Personnel, Product and Environment Protection
HEPA filters (not for chemical vapours)

9. Fume Hoods Removes toxic chemical (ducting sys./ductless)
No HEPA filter -> not for biohazard agents

10. Class 1 BSC Only operator protection (not product)
Biosafety level 1, 2, 3
Inflow away from operator
HEPA filtered exhaust
to environment
Current trend: to Class 2
During review of classifications explain european first (This is sort of like a review of the introduction), then go on to the more detailed american classification
Mention example of british cabinet double exhaust
Also Italian / Australian cabinet style double fan
Also remember to draw out all the airflow in great detail

11. Exhaust Airflow approx. 30%
Class 2 A2
Exhaust Airflow approx. 30%
Recirculating cabinet airflow
No chemical / toxic vapours containment without ducting
Approx. 70% recirculating and 30% exhaust
Downflow approx.
70%
Room Air

12. Class II Type A2 BSC: Ducting
Class II Type A2 can’t be used for chemical vapours. Chemical vapour buildup in cabinet & lab is dangerous. Class II Type A2 BSC can be ducted using a non-airtight (thimble) duct.
During review of classifications explain european first (This is sort of like a review of the introduction), then go on to the more detailed american classification
Mention example of british cabinet double exhaust
Also Italian / Australian cabinet style double fan
Also remember to draw out all the airflow in great detail
Thimble duct: have holes for room air
Bldg. exhaust fluctuations affect cabinet airflow

13. Class II B2 BSC Airflow

14. Class 2 Type B2 Total exhaust cabinet (no recirculation)
Both biological and chemical vapor containment
Care with chemical: should not destroy filter
High exhaust air volume
Interlock system: if building exhaust fails
Dedicated exhaust fan with dynamic balancing
Exhaust fan -> precisely match the cabinet:
- airflow volume
- static pressure
Inflow and downflow are opposite each other
High level of expertise for install & maintenance
During review of classifications explain european first (This is sort of like a review of the introduction), then go on to the more detailed american classification
Mention example of british cabinet double exhaust
Also Italian / Australian cabinet style double fan
Also remember to draw out all the airflow in great detail

15. Class 2 Type B2 BSC: Airflow
During review of classifications explain european first (This is sort of like a review of the introduction), then go on to the more detailed american classification
Mention example of british cabinet double exhaust
Also Italian / Australian cabinet style double fan
Also remember to draw out all the airflow in great detail

16. Class 3 BSC
During review of classifications explain european first (This is sort of like a review of the introduction), then go on to the more detailed american classification
Mention example of british cabinet double exhaust
Also Italian / Australian cabinet style double fan
Also remember to draw out all the airflow in great detail

17. International Standards for Class II
US Standard ANSI/NSF49
European Standard EN12469
Japanese Industrial Standard JIS K3800
South African Standard SABS VC 8041:2001
British Standard BS5726*
German Standard DIN12950 Teil 10*
French Standard NF X44-201:1984*
*now obsolete. Replaced with the harmonized EN12469

18. Biosafety Cabinet Certification

19. Biosafety Cabinet Certification
The functional operation and integrity of each BSC should be certified to national or international performance standards at the time of installation and regularly thereafter by qualified technicians
Evaluation of the effectiveness of cabinet containment should include tests for cabinet integrity, HEPA filter leaks, inflow/downflow velocity profile, air flow smoke pattern, and alarms and interlocks.

20. Testing List

21. Common User Mistakes

22. Common User Mistakes
Confusing a vertical laminar flow cabinet for a Class II BSC
Failure to identify the type of BSC needed for their operations
Class II B2 cabinet not necessarily “safer” than a Class II A2 cabinet (A2 cabinets can also be ducted out when necessary)
Inappropriate choice of installation site / cabinet location
Inappropriate usage / maintenance of the BSC

23. Common User Mistakes: Illustration
Blocking of airflow perforations with objects

24. Proper Installation / Location
Exhaust filter area: Especially susceptible to disruptive air currents. Clearance of 40 cm (minimum) is recommended between the highest point of the cabinet and the ceiling.
Barrier Airflow
Air Inlet
Biosafety Cabinet
Return Air Grille
Personnel Traffic

25. Proper Operation
Slow deliberate movements that will not disrupt airflow, minimize arm movement
When an alarm is activated, do NOT use the cabinet
After usage, wipe down the cabinet with cleaning agents
Work as far into the cabinet as possible
Work starting from clean to “dirty” objects
Do not block airflow perforations with objects/equipments

26. Annual Testing Recommended recertification for BSC’s:
Upon installation at customer’s site
On-site annually in accordance with international standards
When the cabinet is moved

27. UV Lamps
Germicidal UV lamps are not substitutes for proper cleaning of BSC workzone
May cause performance degradation
May compromise personnel safety when proper precautions are not taken
Minimum Intensity is 50 microwatts per square centimeter.

28. Bunsen Burners The use of Bunsen burners in LFC and BSC is discouraged
Compromises cabinet’s operator and cross- contamination protection when used
Filter damage due to bunsen burner usage within workzone

29. Reporting of accident/incident
If there is a spillage in the BSC, notify the PI(s)/supervisor(s)/laboratory officer(s) immediately and decontaminate all objects in the cabinet without turning off the cabinet.

30. Demonstration