The Kinyoun method, or Kinyoun stain (cold method), developed by Robert Koch in 1882 and later modified by other scientist, is an acid-fast procedure used to stain any species of the genus Mycobacterium , Nocardiaand Cryptosporidium species. Certain species of bacteria have a waxy lipid called mycolic acid, in their cell walls which allow them to be stained with Acid-Fast better then a Gram-Stain. The unique ability of mycobacteria to resist decolorization byacid-alcohol is why they are termed acid-fast. It involves the application of a primary stain (basic fuchsin), a decolorizer (acid-alcohol), and a counterstain (methylene blue). Unlike the Ziehl-Neelsen stain (Z-N stain), the Kinyoun method of staining does not require heating. In the Ziehl-Neelsen stain, heat acts as a physical mordant while phenol (carbol of carbol fuschin) acts as the chemical mordant. Since the Kinyoun stain is a cold method (no heat applied), the concentration of carbol fuschin used is increased.
Mycobacterium is a genus of Actinobacteria, given its own family, the Mycobacteriaceae. Over 190 species are recognized in this genus. This genus includes pathogens known to cause serious diseases in mammals, including tuberculosis and leprosy in humans. The Greek prefix myco- means "fungus," alluding to the way mycobacteria have been observed to grow in a mold-like fashion on the surface of cultures. It is acid fast and cannot be stained by the Gram stain procedure.
Nocardia is a genus of weakly staining Gram-positive, catalase-positive, rod-shaped bacteria. It forms partially acid-fast beaded branching filaments. It contains a total of 85 species. Some species are nonpathogenic, while others are responsible for nocardiosis. Nocardia species are found worldwide in soil rich in organic matter. In addition, they are oral microflora found in healthy gingiva, as well as periodontal pockets. Most Nocardia infections are acquired by inhalation of the bacteria or through traumatic introduction.
Cryptosporidium is a genus of apicomplexan parasitic alveolates that can cause a respiratory and gastrointestinal illness (cryptosporidiosis) that primarily involves watery diarrhea with or without a persistent cough in both immunocompetent and immunodeficient humans.
|Application of||Reagent||Cell Colour|
|Acid Fast||Non-acid fast|
|Primary dye||Carbol fuchsin||Red||Red|
|Counter Stain||Methylene blue||Red||Blue|
The Kinyoun method can be modified as a weak acid fast stain, which uses 5% sulfuric acid instead of hydrochloric acid. The weak acid fast stain in addition to staining mycobacteria will stain organisms that are not able to maintain the carbol fuchsin after decolorizing with HCl.
Gram stain or Gram staining, also called Gram's method, is a method of staining used to distinguish and classify bacterial species into two large groups. The name comes from the Danish bacteriologist Hans Christian Gram, who developed the technique.
Staining is a technique used to enhance contrast in samples, generally at the microscopic level. Stains and dyes are frequently used in histology and in the medical fields of histopathology, hematology, and cytopathology that focus on the study and diagnoses disease at a microscopic level. Stains may be used to define biological tissues, cell populations (classifying different blood cells, or organelles within individual cells.
The cell envelope comprises the inner cell membrane and the cell wall of a bacterium. In gram-negative bacteria an outer membrane is also included. This envelope is not present in the Mollicutes where the cell wall is absent.
Acid-fast stain, first introduced by Dr. Paul Ehrlich, also known as the Ziehl–Neelsen staining, is a bacteriological stain used to identify acid-fast organisms, mainly Mycobacteria. It is named for two German doctors who modified it: the bacteriologist Franz Ziehl (1859–1926) and the pathologist Friedrich Neelsen (1854–1898).
The auramine–rhodamine stain (AR), also known as the Truant auramine–rhodamine stain, is a histological technique used to visualize acid-fast bacilli using fluorescence microscopy, notably species in the Mycobacterium genus. Acid-fast organisms display a reddish-yellow fluorescence. Although the auramine–rhodamine stain is not as specific for acid-fast organisms as the Ziehl–Neelsen stain, it is more affordable and more sensitive, therefore it is often utilized as a screening tool.
Acid-fastness is a physical property of certain bacterial and eukaryotic cells, as well as some sub-cellular structures, specifically their resistance to decolorization by acids during laboratory staining procedures. 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.
Fuchsine (sometimes spelled fuchsin) or rosaniline hydrochloride is a magenta dye with chemical formula C20H19N3·HCl. There are other similar chemical formulations of products sold as fuchsine, and several dozen other synonyms of this molecule.
New fuchsine (from German "fuchs", fox) is an organic compound with the formula [(CH3N(H)CH3C6H3)3C]Cl. It is a green-colored solid that is used as a dye of the triarylmethane class. It is one of the four components of basic fuchsine, and one of the two that are available as single dyes. The other is pararosaniline.
Mycobacterium smegmatis is an acid-fast bacterial species in the phylum Actinobacteria 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. It was first reported in November 1884 by Lustgarten, who found a bacillus with the staining appearance of tubercle bacilli in syphilitic chancres. Subsequent to this, Alvarez and Tavel found organisms similar to that described by Lustgarten also in normal genital secretions (smegma). This organism was later named M. smegmatis.
Dr. Franz Ziehl was a German bacteriologist. He was a professor in Lübeck.
Carbol fuchsin, carbol-fuchsin, or carbolfuchsin, is a mixture of phenol and basic fuchsin, used in bacterial staining procedures. It is commonly used in the staining of mycobacteria as it has an affinity for the mycolic acids found in their cell membranes.
Hematoxylin and eosin stain or haematoxylin and eosin stain is one of the principal tissue stains used in histology. It is the most widely used stain in medical diagnosis and is often the gold standard; for example, when a pathologist looks at a biopsy of a suspected cancer, the histological section is likely to be stained with H&E.
Nigrosin is a mixture of synthetic black dyes made by heating a mixture of nitrobenzene, aniline, and hydrochloric acid in the presence of copper or iron. Related to induline, it is a mixture of phenazine-based compounds. Its main industrial uses are as a colorant for lacquers and varnishes and in marker-pen inks. Sulfonation of nigrosin yields a water-soluble anionic dye, nigrosin WS.
Auramine phenol stain is a stain used in clinical microbiology and histology to identify tuberculosis mycobacteria.
The Löwenstein–Jensen medium, more commonly known as LJ medium, is a growth medium specially used for culture of Mycobacterium species, notably Mycobacterium tuberculosis.
Friedrich Carl Adolf Neelsen was a German pathologist.
The Schaeffer–Fulton stain is a technique designed to isolate endospores by staining any present endospores green, and any other bacterial bodies red. The primary stain is malachite green, and the counterstain is safranin, which dyes any other bacterial bodies red.
Moeller staining involves the use of a steamed dye reagent in order to increase the stainability of endospores; carbol fuchsin is the primary stain used in this method. Endospores are stained red, while the counterstain methylene blue stains the vegetative bacteria blue.
Endospore Staining is a technique used in bacteriology to identify the presence of endospores in a bacterial sample, which can be useful for classifying bacteria. Within bacteria, endospores are protective structures used to survive extreme conditions, but this protective nature makes them difficult to stain using normal techniques such as simple staining and Gram staining. Special techniques for endospore staining include the Schaeffer–Fulton stain and the Moeller stain.
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