Putt
for Acid Fast Bacteria

Solutions

Putt's Carbol new fuchsin

Methylene blue, 0.5% in absolute ethanol

Glacial acetic acid, 5% in absolute ethanol

Lithium carbonate, saturated aqueous

Tissue sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are usually satisfactory.

Method

1. Bring sections to water via xylene and ethanol.
2. Immerse in a Coplin jar of carbol new fuchsin for 3 minutes at room temperature.
3. Immerse sections in lithium carbonate for 1 minute and agitate gently.
4. Decolorise with acetic ethanol until sections are pale pink (about 3-5 minutes).
5. Rinse twice with absolute ethanol, 2 minutes each.
6. Counterstain with methylene blue for 1 minute.
7. Absolute ethanol, two changes of 30 seconds each.
8. Clear with xylene and mount with a resinous medium.

Expected results

• Acid fast organisms – red
• Background – blue

Notes

1. Discard the lithium carbonate solution when it turns blue.
2. This method is stated to be suitable for M. leprae.

 

Reference
Putt, F.A., (1972)
Manual of histopathological staining methods
John Wiley & Sons, New York, NY, USA

Translate in Google Translate Instructions