Here is the protocol for acid fast bacilli (AFB).  At the bottom of the post is the link for printing.

Control Tissue : Tissue with AFB present

1.  De-paraffin slides in xylene (1) ——————- 2 minutes (re-use)
2.   De-paraffin slides in xylene (2)——————- 2 minutes (re-use)
3.   Clear slides in 100% alcohol———————- 2 minutes
4.   Clear slides in 100% alcohol———————- 2 minutes
5.   Hydrate slides in 95% alcohol——————– 2 minutes
6.   Hydrate slides in running water—————— 5 minutes
7.   Carbol Fuchsin————————————– 45 minutes (re-use)
8.   Wash in running water—————————– 1 minute
9.   Decolorize in 1% acid alcohol——————— until sections are pale pink
10.   Wash in running water—————————– 8 minutes
11.   Counterstain with working methylene blue—–1 dip (re-use (good for 5 days))
12.   Rinse quickly—————————————- 30 seconds
13.   Dehydrate in 95% alcohol———————— 1 minute
14.   Dehydrate in 100% alcohol———————- 1 minute
15.   Dehydrate in 100% alcohol———————- 2 minutes
16.   Clear in xylene (3)——————————— 2 minutes (re-use)
17.   Clear in xylene (4)——————————— 5 minutes (re-use)

Results:

Acid-fast bacilli —————- Bright Red

Erythrocytes——————– Yellowish Orange

Other tissue elements——- Pale blue

Acid Fast Bacilli