SPORE STAINING PROCEDURE OF BACTERIA EXAMINATION

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SPORE STAINING

This staining is specifically develop for the process of microscopic examination of spore structure of bacteria. The spore wall is shown as impermeable to stain, as a result of that prevent decolourisation by alcohol. The spore are stained by heating the preparation. This process has solution 1 to 4

1 Fuchin-methyl blue stain

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2 Ferric chloride 30% α solutions

3 Sodium sulphate 5%aq solutions

4 Methylene blue 1%aq solutions

 

PROCEDURE

1 Prepare a thin smear film allowed to dry, fix by with small amount of heat.

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2 Apply solutions 1 for 3-5 minute, heat the preparation till steam rises.

3 Wash with water.

4 Apply solutions 2 for 1-2 minute.

5 Replace with solution 3 for 30 second.

6 Wash with water.

7 Apply solutions 4 for 1 minute.

8 Wash and blot carefully, dry with small amount of heat.

Examine microscopically.

Result; spore bright red, the remaining organism stained blue.

Fleming also is a method staining used for examination of bacteria spore.

 

SOLUTIONS

1 Ziehl Neelseen carbol-fuchsin

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2 Decolourizer, this have solution a and b,

(a) Nigrosin 1% solution

(b) Sodium sulphate 5% solutions

3 Nigrosin 10% solutions

basteria spore
BACTERIA SPORE UNDER MICROSCOPE

PROCEDURE

1 Prepare a thin smear film, allowed to dry by little heat.

2 Apply solution 1 for 5 minute heat the preparation until steam rise.

3 Wash with water.

4 Apply solutions 2a for 5-10 second

5 Apply solutions 2b for 5-30 second

6 Wash with water, blot carefully and dry by little heating

7 Drop small volume of solution 3 at one end of the slide and spread it on the surface of the stained preparation by using another glass slide.

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8 Allowed to dry and examine with microscope.

Result; spore stain bright red other organism unstained against dark grey background of nitrosin.

 

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