Plant tissue culture is a sterile laboratory method for growing plant cells, tissues, or organs on nutrient-rich culture media. This process, also known as micropropagation, leverages the plant cells' totipotency to produce thousands of genetically identical plant clones, or soma clones. Key advantages include rapid, large-scale plant production, the creation of disease-free plants from infected parent stock using meristems, the conservation of endangered species, and a role in plant biotechnology and research. The Process Explained Explant Selection: A small piece of healthy plant tissue, called an explant, is taken from a parent plant. Sterilization: The explant is thoroughly sterilized to remove any potential contaminants like bacteria or fungi that could interfere with the growth process. Culture

1. Welcome You

2. MAHATMA PHULE KRISHI VIDYAPEETH , RAHURI
COLLEGE OF AGRICULTURE, LONI
2020-2021
Under the Department
Of Agricultural Botany
SRP-EL BOT- 408
By
Shinde Amruta Balasaheb
(AGL/53/2018)
Module In charge
Prof. G.B. Labade
College of Agriculture, Loni
Sterilization Techniques & Handling of
Equipments

3. Defination of Sterilization :
Sterilization is a process of
destruction of all forms of living microorganisms from a
substance.

4. METHODS OF STERILIZATION
• THREE METHOD:-
Chemical
Mechanical
Physical

5. PHYSICAL METHOD
1)Sunlight
2)Drying
3)Heat:- a) Dry heat
b) Moist heat
4)Radiation

6. 1) Sunlight –
 It is responsible for spontaneous sterilization in
natural conditions .
By killing bacteria suspended in water , sunlight
provides natural method of disinfection of tanks &
lakes.
2) Drying –
Moisture is essential for growth of bacteria .
Drying in air has dangerous effect on many
bacteria .
However, Viruses & spores are unaffected.

7. a) Dry Heat :
 In dry heat sterilization, dry heat used for sterilizing
different materials.
 The temp. required is 160 c for 1 hour.
⁰
 It is suitable method for sterilization of substances
destroyed by moisture.
3) Heat sterilization: -
Its has two types
b) Moist Heat :
 In moist heat sterilization, Heating process
in autoclave .
 The recommendation temp. in autoclave is
121 c for 15 min.
⁰

8. Advantages of Physical Sterilization:-
1) Effective method of sterilization.
2) Articles remain dry after sterilization.
Disadvantages :-
3) Drenching & wetting articles may occurs.
4) Takes longer time compared to autoclave.

9. Chemical Methods
1 ) Gaseous sterilization:-
a) Ethylene oxide
b) Formaldehyde gas
2 ) Liquid sterilization :-
a) Alcohol
b) Phenol

10. 1. Gaseous sterilization:-
a ) Ethylene oxide :-
 Easily dissolved in water & organic solvent.
Effective against all types of microorganisms including
viruses & spores .
The gas used is safe & non-inflammable.

11. B. Formaldehyde gas:-
It is a simple chemical compound made up of
hydrogen, oxygen and carbon.
High level disinfectant.
For better effect it needs relative humidity of 60-
80% and temperature of 37⁰c .

12. 2. Liquid sterilization
a. Alcohols
Alcohols are effective disinfectants for many reasons.
They evaporate quickly, without leaving a residue.
They are used mainly to disinfect external surfaces of
equipments.
b. Phenols
Phenol is one of the oldest antiseptic agents.
It acts by damaging cell membrane.
Phenols are used for decontamination of laboratory
surface nono critical medical items.

13. Advantages of Chemical Sterilization
1 . Technically easier.
2 . Suitable for equipments damaged by heat sterilization.
Disadvantages
1 . Act only on exposed surfaces.
2 . Some chemicals react with metals.

14. 3.FILTRATION (MECHANICAL) METHOD:-
 Filtration sterilization used for heat sensitive
materials to sterilize.
 Filtration process does not destroy but remove the
microorganisms.
 Sterilization test must be done.

15. Advantages of Mechanical
Sterilization:-
1. Filtration is quite rapid.
2. Bacteria are removed by sieving.
Disadvantages:-
3. Chemically less resistance.
4. Soluble in certain organic solvents.

16. Instruments Used:-
1 ) Hot air oven
2) Autoclave
3) Laminar air flow

17. Hot Air
Oven
Hot air ovens are electrical
devices which use dry heat to
sterilize .
An oven provides a temperature
higher than that of atmosphere.
The temperature range covered
by oven is between 50-250⁰C.
Use:-
Used for rapid evaporation
of materials, rapid drying & for
sterilization of articles that can be
sterilized by dry heat.

18. Autoclave
An autoclave is a machine that
provides a physical methods of
sterilization by killing bacteria
The recommendation
temperature for sterilization in
autoclave is 121-124⁰C.
Use:-
To sterilize the culture
media.

19. Laminar Air
Flow
Definition:-
Laminar air flow is an
enclosed bench designed to prevent
contaminations like biologicals
particles or any particle sensitive
device.
Use:-
Laminar Air Flow provides
a work area with aseptic
conditions for the tissue culture.

20. THANK YOU