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Endospore staining is a technique used in bacteriology to identify the presence of endospores in a bacterial sample. [1] Within bacteria, endospores are protective structures used to survive extreme conditions, including high temperatures making them highly resistant to chemicals. [ 2 ]
The Schaeffer–Fulton stain is a technique designed to isolate endospores by staining any present endospores green, and any other bacterial bodies red. [1] The primary stain is malachite green , and the counterstain is safranin , which dyes any other bacterial bodies red.
Safranin (Safranin O or basic red 2) is a biological stain used in histology and cytology. Safranin is used as a counterstain in some staining protocols, colouring cell nuclei red. This is the classic counterstain in both Gram stains and endospore staining. It can also be used for the detection of cartilage, [2] mucin and mast cell granules.
Endospore staining is particularly useful for identifying endospore-forming bacterial pathogens such as Clostridioides difficile. Prior to the development of more efficient methods, this stain was performed using the Wirtz method with heat fixation and counterstain.
A counterstain is a stain with colour contrasting to the principal stain, making the stained structure easily visible using a microscope. Examples include the malachite green counterstain to the fuchsine stain in the Gimenez staining technique and the eosin counterstain to haematoxylin in the H&E stain . [ 1 ]
In the Gimenez staining method, basic fuchsin stains bacteria red or magenta, and malachite green is used as a blue-green counterstain. Malachite green is also used in endospore staining, since it can directly stain endospores within bacterial cells; here a safranin counterstain is often used. Malachite green is a part of Alexander's pollen stain.
Endospore made visible with a stain. 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.
Fungal yeast forms are inconsistently stained with Acid-fast stain which is considered a narrow spectrum stain for fungi. [21] In a study on acid-fastness of fungi, [ 22 ] 60% of blastomyces and 47% of histoplasma showed positive cytoplasmic staining of the yeast-like cells, and Cryptococcus or candida did not stain, and very rare staining was ...