Kinyoun stain

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The Kinyoun stain is a method of staining acid-fast microorganisms, specifically Mycobacterium and Nocardia. The procedure for Kinyoun staining is similar to the Ziehl-Neelsen stain, but does not involve heating the slides being stained.1 The Kinyoun staining method uses carbol fuchsin as a primary stain, followed by decolorization with an acid-alcohol solution and methylene blue as a counterstain. Kinyoun carbol fuschsin has a greater concentration of phenol and basic fuchsin and does not require heating in order to stain properly.1 When viewed under a microscope, a Kinyoun stained slide will show acid-fast organisms as red and nonacid-fast organisms as blue.

Procedure

  1. Flood slides with Kinyoun carbol fuchsin for 5 minutes.
  2. Rinse gently with water until the water flows off clear.
  3. Flood slides with acid-alcohol (3% HCl in ethanol) for 3~5 seconds.
  4. Rinse gently with water until the water flows off clear.
  5. Flood slides with methylene blue for 3 minutes.
  6. Rinse gently with water until the water flows off clear.
  7. Allow slides to air dry before viewing.

Modifications

A solution of 1% sulfuric acid can be substituted in place of 3% HCl solution. The sulfuric acid solution does not decolorize as strongly as the hydrochloric acid. This makes it useful for staining organisms that are weakly acid fast, such as Nocardia. Brilliant Green may be substituted for Methylene Blue as a counterstain, resulting in non-acid fast organisms appearing green rather than blue.

Another alternative is 20% sulphuric acid instead of HCl.

See also

Bibliography

  • Bartelt, Margaret (October 4, 1999). Diagnostic Bacteriology: A Study Guide (in English) (First ed.). F.A. Davis Company. p. 500. ASIN 0803603010. ISBN 978-0803603011. 
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