Acid Fast Bacilli (AFB) Protocol

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

Human Kidney AFB
Human Kidney AFB
  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


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