Explain Procedure for Acid Fast Staining?

Carry out the exercise using the steps enumerated herewith.

1. Label the clean, non-greasy slides and make smears of M. smegmatis, S. aureus and mixture of two on them.

2. Allow the smear to air dry and then fixed the smear by heating.

3. Cover the smear with carbol fuchsin and put the slide on a beaker of warm water. Allow the steam to pass through smear for five minutes. Replenish the stain, if needed.

4. Allow the slide to cool. Then wash the smear with tap water.

5. Decolourize the smear by adding acid alcohol drop wise till the clean alcohol with slight red tinge comes out.

6. Wash the slide with tap water.

7. Flood the smear with methylene blue for two minutes.

8. Wash the smear with tap water. 9. See under 100x after air drying.