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Acid-fast Ziehl-Neelsen Staining of Bacteria - P2
Ziehl's carbol fuchsin stain being applied to slide over a heated water bath.
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Steps of Acid Fast Staining Procedure
The protocol for staining acid-fast organisms is as follows (see photos below):
- Place strip of blotting paper over the slide.
- Place slide over a screened water bath. Then saturate blotting paper with primary stain Ziehl’s carbol fuschion.
- Allow slide to sit over water bath for 3–5 minutes. Reapply stain if it begins to dry out.
- Discard blotting paper. Rinse slide until water runs clear.
- Flood slide with decolorizer, Acid Alcohol, for 10–15 seconds. Then rinse.
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SPO VIDEO: How to Do an
Acid-fast Bacterial Stain
- Flood slide with counterstain, Crystal Violet, for one minute. Then rinse.
- Gently blot the slide dry. It is now ready to be viewed under oil immersion (1000x TM) with a compound light microscope.
