Protocol
Adams
for Acid Alcohol Fast Bacteria
Materials
- Goodpasture’s carbol–fuchsin
- Weigert’s iron hematoxylin (1904)
- 0.1% hydrochloric acid in 50% ethanol
- 5% aqueous acetic acid
- 0.1% light green SF yellowish in 0.2% acetic acid
Tissue Sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are usually satisfactory.
Protocol
- Bring sections to water via xylene and ethanol.
- Stain nuclei with Weigert’s iron hematoxylin for 1 minute.
- Rinse well with 0.1% hydrochloric acid in 50% ethanol.
- Repeat the following 2 steps four times:
- Place in Goodpasture’s carbol fuchsin for 1 minute.
- Decolorize with 5% acetic acid until no more color is extracted.
- Wash with water.
- Counterstain with light green solution for 1 minute.
- Dehydrate with 85% and absolute ethanols.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – black
- Acid fast organisms – red
- Background – green
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Gray, Peter. (1954)
The Microtomist’s Formulary and Guide.
Originally published by: The Blakiston Co.
Republished by: Robert E. Krieger Publishing Co.