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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.
An endospore stain of the cell Bacillus subtilis showing endospores as green and the vegetative cell as red Phase-bright endospores of Paenibacillus alvei imaged with phase-contrast microscopy. An endospore is a dormant, tough, and non-reproductive structure produced by some bacteria in the phylum Bacillota.
Differential staining uses multiple stains per slide. Based on the stains being used, organisms with different properties will appear different colors allowing for categorization of multiple specimens. Differential staining can also be used to color different organelles within one organism which can be seen in endospore staining. [1]
Another endospore stain of B. subtilis Bacillus subtilis can divide symmetrically to make two daughter cells (binary fission), or asymmetrically, producing a single endospore that can remain viable for decades and is resistant to unfavourable environmental conditions such as drought , salinity , extreme pH , radiation , and solvents .
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.
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.
Fig1. The sporulation process of Bacillus subtilis. Bacillus subtilis is a rod-shaped, Gram-positive bacteria that is naturally found in soil and vegetation, and is known for its ability to form a small, tough, protective and metabolically dormant endospore.