Here's a summary of the acid-fast stains used for Mycobacteria:

Ziehl-Neelsen (ZN) Stain

1. Primary stain: Carbol fuchsin (red)
2. Decolorizer: Acid-alcohol (3% HCl in 95% ethanol)
3. Counterstain: Methylene blue (blue)
4. Result: Mycobacteria appear red, background appears blue

Kinyoun Stain

1. Primary stain: Carbol fuchsin (red)
2. Decolorizer: Acid-alcohol (1% HCl in 95% ethanol)
3. Counterstain: Methylene blue (blue)
4. Result: Mycobacteria appear red, background appears blue

Auramine-Rhodamine Stain

1. Primary stain: Auramine O (yellow) and Rhodamine B (pink)
2. Decolorizer: Acid-alcohol (0.5% HCl in 95% ethanol)
3. Counterstain: Potassium permanganate (colorless)
4. Result: Mycobacteria appear yellow or pink, background appears dark

All three stains are used to detect acid-fast bacteria, including Mycobacteria. The ZN stain is the most commonly used, while the Kinyoun stain is a modification of the ZN stain. The Auramine-Rhodamine stain is a fluorescent stain, requiring a microscope with a fluorescent light source.

These stains work by:

1. Penetrating the Mycobacterial cell wall with the primary stain
2. Resisting decolorization with acid-alcohol due to the unique cell wall composition
3. Appearing colored against a contrasting background

This characteristic acid-fastness helps identify Mycobacteria and differentiate them from other bacteria.


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