SPORE STAINING PROCEDURE OF BACTERIA EXAMINATION

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

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.

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

2 Decolourizer, this have solution a and b,

(a) Nigrosin 1% solution

(b) Sodium sulphate 5% solutions

3 Nigrosin 10% solutions

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.

8 Allowed to dry and examine with microscope.

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