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ACID FAST Os Contains waxlike lipoidal material affecting staining quality. Carbolfuchsin is primary stain. Acid fast organisms resist decolorization with acid alcohol. After decolorization, methyelene blue is added to organisms to counterstain any material that is not acid fast. It is used for examination of Mycobacterium.
The principle of staining is depend on the resistance of this type of bacteria to decolarization by acid alcohol, because the cell wall contain waxy material (mycolic acid) which prevent the removal of carbol fuchsin from the cell.
Staining Procedure
1. Flood the slide with strong carbol fuchsin and heat until steam rises (but do not boil).
2. After 3-4 min apply more heat until rises again; do not let the stain dry on the slide.
3. About 5-7 min after the first application of heat washes the slide thoroughly under running water.
4. Decolourize in acid-alcohol until all traces of red have disappeared from the film. Decolourization should not be attempted in one stage; there should be intermittent washings in water and re-application of acid-alcohol.
5. Wash well with water and counter stain with Methylene blue for 1 min.
6. Wash and stand end to drain.
7. Acid-fast organisms are red, other organisms are blue.
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Iodine is used to bind the Crystal Violet to the Gram Positive microbes.
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What is the action of iodine in grams stain?
Iodine is used to bind the Crystal Violet to the Gram Positive microbes.
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