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Ziehl–Neelsen stain (classic and modified bleach types) [5]; Kinyoun stain; For color blind people (or in backgrounds where detecting red bacteria is difficult), Victoria blue can be substituted for carbol fuchsin and picric acid can be used as the counter stain instead of methylene blue, and the rest of the Kinyoun technique can be used.
The Ziehl-Neelsen stain, also known as the acid-fast stain, is a bacteriological staining technique used in cytopathology and microbiology to identify acid-fast bacteria under microscopy, particularly members of the Mycobacterium genus.
The Kinyoun method or Kinyoun stain (cold method), developed by Joseph J. Kinyoun, is a procedure used to stain acid-fast species of the bacterial genus Mycobacterium. [1] It is a variation of a method developed by Robert Koch in 1882.
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.
[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]
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.
The two most common methods for visualizing these acid-fast bacilli as bright red against a blue background are the Ziehl-Neelsen stain and modified Kinyoun stain. Fite's stain is used to color M. leprae cells as pink against a blue background. Rapid Modified Auramine O Fluorescent staining has specific binding to slowly-growing mycobacteria ...
The high mycolic acid content of Mycobacteria, is responsible for the staining pattern of poor absorption followed by high retention. The most common staining technique used to identify acid-fast bacteria is the Ziehl–Neelsen stain or acid-fast stain, in which the acid fast bacilli are stained bright red and stand out clearly against a blue ...