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The Ziehl-Neelsen stain is a two step staining process. In the first step, the tissue is stained with a basic fuchsin solution, which stains all cells pink. In the second step, the tissue is incubated in an acid alcohol solution, which decolorizes all cells except for acid-fast cells, which retain the color and appeared as red.
English: This is a diagram of the basic steps of a Ziehl-Neelsen (Acid Fast) staining procedure File:Basic steps of acid fast staining procedure.svg is a vector version of this file. It should be used in place of this PDF file when not inferior.
[1] [2] Once stained as part of a sample, these organisms can resist the acid and/or ethanol-based decolorization procedures common in many staining protocols, hence the name acid-fast. [ 2 ] The mechanisms of acid-fastness vary by species although the most well-known example is in the genus Mycobacterium , which includes the species ...
The Kinyoun method can be modified as a weak acid fast stain, which uses 0.5–1.0% sulfuric acid instead of hydrochloric acid.The weak acid fast stain, in addition to staining Mycobacteria, will also stain organisms that are not able to maintain the carbol fuchsin after decolorizing with HCl, such as Nocardia species and Cryptosporidium.
[2] [3] Carbol fuchsin is used as the primary stain dye to detect acid-fast bacteria because it is more soluble in the cells' wall lipids than in the acid alcohol. If the bacteria is acid-fast the bacteria will retain the initial red color of the dye because they are able to resist the destaining by acid alcohol (0.4–1% HCl in 70% EtOH). [4]
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A Ziehl–Neelsen stain is an acid-fast stain used to stain species of Mycobacterium tuberculosis that do not stain with the standard laboratory staining procedures such as Gram staining. This stain is performed through the use of both red coloured carbol fuchsin that stains the bacteria and a counter stain such as methylene blue.
Mycobacterium smegmatis is an acid-fast bacterial species in the phylum Actinomycetota and the genus Mycobacterium.It is 3.0 to 5.0 μm long with a bacillus shape and can be stained by Ziehl–Neelsen method and the auramine-rhodamine fluorescent method.