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The decolorization step is critical and must be timed correctly; the crystal violet stain is removed from both gram-positive and negative cells if the decolorizing agent is left on too long (a matter of seconds). [19] After decolorization, the gram-positive cell remains purple and the gram-negative cell loses its purple color. [19]
Gram-positive bacteria retain the crystal violet stain used in the test, resulting in a purple color when observed through an optical microscope. The thick layer of peptidoglycan in the bacterial cell wall retains the stain after it has been fixed in place by iodine. During the decolorization step, the decolorizer removes crystal violet from ...
The stain proved popular and in 1884 was used by Hans Christian Gram to stain bacteria. He credited Paul Ehrlich for the aniline-gentian violet mixture. [ 32 ] Grübler's gentian violet was probably very similar, if not identical, to Lauth's methyl violet, which had been used as a stain by Victor André Cornil in 1875.
Schematic of typical Gram-positive cell wall showing arrangement of N-Acetylglucosamine and N-Acetylmuramic acid; Teichoic acids not shown.. The Gram-positive cell wall is characterized by the presence of a very thick peptidoglycan layer, which is responsible for the retention of the crystal violet dyes during the Gram staining procedure.
Gram-positive bacteria have a thick peptidoglycan layer in their cell wall, which retains the crystal violet during Gram staining, resulting in a purple color. Gram-negative bacteria have a thin peptidoglycan layer which does not retain the crystal violet, so when safranin is added during the process, they stain red.
The negatively charged cell wall of many microorganisms attracts the positively charged chromophore which causes the specimen to absorb the stain giving it the color of the stain being used. Positive staining is more commonly used than negative staining in microbiology. The different types of positive staining are listed below. [1]
The gram-positive cocci are a large group of bacteria with similar morphology. All are spherical or nearly so, but they vary considerably in size. Members of some genera are identifiable by the way cells are attached to one another: in pockets, in chains, or grape-like clusters.
Main staining types when using hematoxylin and eosin (H&E). A Basophil granulocyte stains dark purple upon H&E staining. Basophilic is a technical term used by pathologists. It describes the appearance of cells, tissues and cellular structures as seen through the microscope after a histological section has been stained with a basic dye.
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