STANDARD OPERATING PROCEDURES FOR ACID FAST STAIN

1. SPECIMEN:

1. Gram stain can be done on various samples such as sputum(avoid saliva), pus, and tissue,etc.

2. REAGENTS:

a. Carbol Fuchsin

b. Acid-alcohol

c. Methylene blue

3. PROCEDURE:

1. Sample is fixed by heat.
2. Flood the slide with carbol fuchsin stain.
3. Heat the slide gently until it steams for about 5 minutes.
4. Pour off the carbol fuchsin and wash slide thoroughly with water.
5. Decolourize with acid-alcohol for about 5 minutes.
6. Wash slide thoroughly with water.
7. Flood slide with methylene blue counterstain for 1 min.
8. Wash with water and blot excess water and dry.
9. Observe the slide under 100X microscope.

a. The acid-fast bacilli are red color rods.

10. Prepare lab report of the observations.