Lab aseptic techniques

1. Aseptic
Techniques.
(Plant tissue culture)

2. What is Aseptic Techniques??
 Aseptic technique refers to a procedure
that is performed under sterile
conditions
 This includes medical and laboratory
techniques which deals with cultures and
human cells and tissue transplantation
 Aseptic is a method used to prevent
contamination with microorganisms

3. What is the aim of Aseptic
Technique???
To prevent the access of micro-organism during the
preparation and testing

4. What is a culture??
A microbiological culture, or
microbial culture, is a method of
multiplying microbial organisms
by letting them reproduce in
predetermined culture medium
under controlled laboratory
conditions.
A microbiological culture, or
microbial culture, is a method of
multiplying microbial organisms by
letting them reproduce in
predetermined culture medium
under controlled laboratory
conditions.

5. • Plant tissue culture is rich
in source of nutrients
• Microbes can grow very fast
on it as compared to plants,
which is very harmful for
plants
• Therefore aseptic
conditions should be
maintained in the culture
bottle

6. Sources of Contamination
Sources of Contamination
1. The Atmosphere.
2. The Breath.
3. The Hands

7. 4. The Clothing
5. The Working Surface
6. The Equipment

8. What need to be sterilized in the process
of aseptic technique??
1. Culture Media
2. Fluids used in the lab
3. Reagents
4. Laboratory containers
5. Laboratory Equipment's

9. Methods of Sterilization
A :- Physical Methods
Dry heat:
i. Hot air oven Glassware
ii. Flaming Inoculation loop and needle
Moist-heat (steam under high pressure)
i. Autoclave Culture Media

10. Filtration
Radiation
Used for heat sensitive
liquids like, Antibiotic
solutions and Sera
Used for enclosed area such
as inoculation room and
virus lab

11. B :- Chemical Methods
Alcohols
Aldehydes
Phenolics
Here the surface, skin etc.
is sterile
eg: ethyl alcohol 70%
conc.
Here equipment such as
Centrifuge is sterile
eg: Formaldehyde
Here floors, walls,
slabs are sterile
Eg: Phenols
(Chloroxylenol)

12. Instruments for Plant Tissue
Culture
These are Sterilized by
dipping in 95% ethanol
followed by flaming and
cooling
needle
Forceps blunt
and sharp
scalpel
rod

13. Environment of the tissue
culture laboratory
 Media room
Maintain cleanliness, removal of contaminated culture, and
restricted entry is done
 Washing room
Contaminated cultured should be autoclaved and discarded
with utmost care and maintain general cleanliness
 Transfer area
A sterile area is required for performing various aseptic
manipulations during tissue culture. This ensures that
contaminants do not gain entry into the culture via Laminar
air flow cabinets of various shapes and sizes are available
commercially. These cabinets allows the tissue culturists to
work in the sterile environment for long stretch of time. It
provides a covered enclosed area for working

14. Laminar air flow cabinet

15.  Clean hand and forearm properly before starting
the work
 Hands may be cleaned with dilute solution of
alcohol or commercial hand sanitizers available in
the market
 Wearing a surgical mask and head gear and lab
coat while working in the laminar will reduce the
chances of contamination
 Presence of other persons in or near the transfer
area should be minimized .
Operator precautions:-

16. >>>Thankuh!!