This presentation can help to gain the knowledge about the pure culture technique method as a spread plate technique. The laboratory uses can helpful to gain knowledge

1. SPREAD PLATE
TECHNIQUE
M.J. AFRA
PG Scholar
Department Of Microbiology
Thassim Beevi Abdul Kader
College For Women
Kilakarai

2. AIM
To perform spread plate technique
To perform isolation of
microorganisms from the sample
To isolate mutagenic or converted
strains of microorganisms by using
differential new products

3. PRINCIPLE
A disadvantage of pour plate method is to overcome
with the help of spread plate method
This method is performed for the assay of chemicals
like antibiotics, vitamins, etc.,
This method is called a spread plate because L-rod
or cotton swab is used to spread the sample.
This technique also used in the isolation and
enumeration of microorganisms from samples with
lower populations of bacteria and other
microorganisms

4. MATERIALS REQUIRED
Test tubes with 9ml of diluents
Cotton
Autoclave
Hot air oven
Water bath
Nutrient agar
Petri plates
L-Rod and Pipettes
Sprit

5. PROCEDURE
1) Prepare nutrient agar medium, sterilize at 121o C and pour it into
petri plates and allow to solidify
2) Dilute the sample upto 10-5
3) Add 0.1ml of sample from 10-3 onto the center of an agar medium
using sterile pipette
4) Perform similar procedure for 10 -4 and 10 -5 dilutions
5) Dip the L-Rod into a beaker of spirit
6) Breafly flame ethanol soaker spreader on Bunsen burner and allow it
to cool
7) Spread the sample evenly on the agar plate’s surface
8) Incubate all the plates at appropriate temperature (Bacteria – 37oC &
Fungus – 25oC to 30oC) for 24 – 48 hours and observe the results

6. ADVANTAGES AND
DISADVANTAGES
ADVANTAGES DISADVANTAGES
Very small quantity sample
is enough to enumerate
the microorganisms
In this methos, only
aerobes and
microaerophiles may be
eliminated

7. RESULTS
Individual colonies are noted at
highest dilution and counted
properly. This technique also used
to perform assay procedures