2. introduction
• What is Laminar Air Flow
• Types of laminar air flow:
• working principle:
• Management of Laminar air flow
• Safety & Precautions taken for Laminar Hood
• validation
3. What is Laminar Air Flow
• Laminar Air Flow is an enclosed bench designed to prevent
contaminations like biological particles or any particle sensitive
device
• This closed cabinet is usually made up of stainless steel without
any gap or joints where spores might collect.
• Laminar Hoods are equipped with a shortwave ultraviolet
germicidal lamp to sterilize the shell
• Use:Laminar Air Flow provides a work area with aseptic/sterile
conditions for the tissue culture
4.
5. Types of laminar air flow:
• . Laminar Flow Cabinets can be produced as both horizontal and
vertical cabinets
• There are many different types of cabinets with a variety of
airflow patterns for different purposes
• Vertical Laminar Flow Cabinets
• Horizontal Laminar Flow Cabinets
• Laminar Flow Cabinets and Hoods
• Laminar Flow Benches and Booths
6. • Horizontal Laminar Flow Cabinets:
• direction of air flow which comes from above
• then changes direction and is processed across the work in a
horizontal direction.
• The constant flow of filtered air provides material and product
protection.
• Vertical Laminar Flow Cabinets
• function equally well as horizontal Laminar Flow Cabinets
• laminar air directed vertically downwards onto the working area
The air can leave the working area via holes in the base
• Vertical flow cabinets can provide greater operator protection.
7. working principle:
• A laminar flow hood consists of a filter pad, a
fan and a HEPA (High Efficiency Particulates
Air) filter
• In a laminar flow hood the air is passed
through a HEPA (High Efficiency Particulates
Air) filter which removes all airborne
contamination to maintain sterile conditions.
• The fan sucks the air through the filter pad
where dust is trapped.
8. working principle:
• After that the prefiltered air has to pass the
HEPA filter where contaminating fungi,
bacteria, dust etc are removed.
• Now the sterile air flows into the working
area where you can do all your flasking work
without risk of contamination.
9. Management of Laminar air flow
• Check and make sure that the area is clean & free from the
previous product and any other materials related to previous
products
• There are three switches and a pressure barometer side panel of
the instrument
• Switch (1) AIR-FLOW
• Switch (2) LIGHT
• Press ’switch (1)’ to start the AIR-FLOW through the HEPA Filters
• The AIR-FLOW should be kept ‘ON’ for about 5 minutes before
carrying out any work under laminar flow.
10. Management of Laminar air flow
•Check the level of red-oil indicator should be at 10 to15 mm mark of water
column, when AIR-FLOW is ‘ON’.
•Switch ‘OFF’ the instrument when not in use.
•UV lamp that should be turned on about 30 minutes before being used to
sterilize the shell or cabinet or the surface of the Laminar Air Flow to avoid any
kind of contaminations
•Wipe down the surface with 95% ethanol before and after each use.
• After inoculation, the cultures were transferred to the culture room which was
initially sterilized by washing all of its walls and floor
•The instruments like foreceps, inoculating neadles, the scalpel blade etc.were
flame sterilized before using them for the inoculation
11. Management of Laminar air flow
• The finger tips were cleaned by 70% ethyl alcohol .
• Mouth and nose were covered with a mask so as to avoid the
contamination of the cultures. Flame Sterilization
• Surface Sterilization of Explant • The explants wear treated with 0.1%
Mercuric Chloride (HgCl2)for 10 minutes Explant
• Preparation of Explant • The explants were further trimmed and extra
outer portain wear removed to make them stable size. Trimming
explant
• . Inoculation of Explant • After cutting of explant were transfer to
culture bottle containing MS media • Innoculated plant then
transfer to culture room Inoculation
12. Safety & Precautions taken for Laminar Hood
• It is important to switch off UV light during use as it may cause
mutations.
• Large objects should never be placed near the back of the hood.
Which normally suspends the contaminants and removes them
from the area.
• Waste and other items should never enter the hood. Hands
should be cleaned by ethanol.
• Do not touch hair, face or clothing while working. Excess dust
should be removed from items before introducing them into the
hood.
• Always handle open bottle at an angle; never let them point
directly up
13. validation
• Validation is "Establishing documented evidence that provides a
high degree of assurance that a specific process will consistently
produce a product meeting its pre-determined specifications and
quality attributes.
• Design qualification(DQ)
• Installation qualifications(IQ)
• Operational qualifications (OQ)
• Performance qualification (PQ)
14. The common requirements outlined in the "General" section are
required. Additionally, product contact surfaces must be stainless
steel or approved plastics or rubber
• Operational Qualification:
• In addition to the common requirements outlined in the "General"
section, the OQ protocol will outline the following tests
15. HEPA Filter Integrity
Test
Photometer readings downstream of the HEPA filter unit will be less than
or equal to 0.01% of the upstream challenge.
Repaired areas are not to exceed 5% of any filter face area.
Biosafety Cabinet
Construction Certification
All welds, gaskets, plenum penetrations and seals will hold pressure within
10% for 30 minutes or be free of soap bubbles after sealed cabinet is
pressurized with compressed air to 2" water gauge.
Air Velocity
Critical Areas should have laminar airflow of 90 fpm ± 20%, or 28 m/min ±
20%
Airflow, Laminar Flow
Hoods
During average supply air velocity testing, the average reading must be
between 120-130 fpm, or 35-40 m/min.
Airflow, Biosafety
Cabinets
The calculated average intake air velocity must be between 100-130 fpm or 30-
40 m/min
16. Performance Qualification
Smoke Testing
Airflow will be unidirectional
During airflow smoke pattern testing, airflow along the entire perimeter of the work
access area will be inward; the smoke will flow downward with no dead spots or
reflux within the cabinet; no airflow escape from the cabinet and no billowing over
or penetrating the work surface
Total Particulate Counts
The area meets the acceptance criteria for total particulates if the average of the particle
concentrations measured at each location falls at or below the class limit.
Any excursion of the class limit for an individual run must be noted, investigated and
justified
Slit to Agar Viable Air
Sampling
less than or equal to 0.1 CFU/ft3
Viable Surface
Sampling
less than or equal to 2 CFU per 30 cm2
Culture media testing Culture media are positive in growth promotion studies.