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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. Safranin typically has the chemical structure shown at right (sometimes described as dimethyl ...
Gram-positive bacteria have a thick mesh-like cell wall made of peptidoglycan (50–90% of cell envelope), and as a result are stained purple by crystal violet, whereas gram-negative bacteria have a thinner layer (10% of cell envelope), so do not retain the purple stain and are counter-stained pink by safranin. There are four basic steps of the ...
Crystal violet stains both Gram positive and Gram negative organisms. Treatment with alcohol removes the crystal violet colour from gram negative organisms only. Safranin as counterstain is used to colour the gram negative organisms that got decolorised by alcohol. While ex vivo, many cells continue to live and metabolize until they are "fixed".
One commonly recognizable use of differential staining is the Gram stain. Gram staining uses two dyes: Crystal violet and Fuchsin or Safranin (the counterstain) to differentiate between Gram-positive bacteria (large Peptidoglycan layer on outer surface of cell) and Gram-negative bacteria. Acid-fast stains are also differential stains.
For both gram-positive and gram-negative bacteria, particles of approximately 2 nm can pass through the peptidoglycan. [7] It is difficult to tell whether an organism is gram-positive or gram-negative using a microscope; Gram staining, created by Hans Christian Gram in 1884, is required. The bacteria are stained with the dyes crystal violet and ...
Malachite green is water-soluble so vegetative cells and spore mother cells can be decolorized with distilled water and counterstained with 0.5% Safranin. [7] In the end, a proper smear would show the endospore as a green dot within either a red or pink-colored cell. [2]
In Gram staining, crystal violet stains only Gram-positive bacteria, and safranin counterstain is applied which stains all cells, allowing the identification of Gram-negative bacteria as well. An alternative method uses dilute carbofluozide.
A number of other bacteria—that are bounded by a single membrane, but stain gram-negative due to either lack of the peptidoglycan layer, as in the mycoplasmas, or their inability to retain the Gram stain because of their cell wall composition—also show close relationship to the gram-positive bacteria.