How use in tissue culture lab

1. Management of Laminar Air Flow and
Inoculation of Explant
Presentated by:
Tanmoy Mondal
Reg.no:H-2014-19-B

2. 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

3. History of Laminar Air Flow
• Laminar Air Flow is a fairly new
innovation.
• The idea of Laminar Air Flow was
develop by few scientist around
early 2000s.
• Johan Coecke et al. gave the
basic definition of Laminar Air
Flow in 2005 Johan Coecke et al.

4. Structure of Laminar air flow
Caster
HEPA filter
Control panel
UV light
Working Table
Pre-Filter

5. Working Principal
• 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.
• 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.

6. Management of Laminar air flow
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

7. Management of Laminar air flow
The instruments like
foreceps, inoculating
neadles, the scalpel blade
etc.were flame sterilized
before using them for the
inoculation.
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

8. Surface Sterilization of Explant
• The explants wear treated
with 0.1% Mercuric
Chloride (HgCl2)for 10
minutes
Explant

9. Preparation of Explant
• The explants were further
trimmed and extra outer
portain wear removed to
make them stable size.
Trimming explant

10. Inoculation of Explant
• After cutting of explant were
transfer to culture bottle
containing MS media
• Innoculated plant then
transfer to culture room
Inoculation

11. Culture Room

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,
 Properly adjust the flame of the Bunsen burner. The proper flame is a
small blue cone; it is not a large plume, nor is it orange.

13. Some Activities