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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]
Unlike negative staining, positive staining uses basic dyes to color the specimen against a bright background. While chromophore is used for both negative and positive staining alike, the type of chromophore used in this technique is a positively charged ion instead of a negative one. The negatively charged cell wall of many microorganisms ...
Both gram-positive and gram-negative bacteria commonly have a surface layer called an S-layer. In gram-positive bacteria, the S-layer is attached to the peptidoglycan layer. Gram-negative bacteria's S-layer is attached directly to the outer membrane. Specific to gram-positive bacteria is the presence of teichoic acids in the cell wall. Some of ...
Teichoic acids give the gram-positive cell wall an overall negative charge due to the presence of phosphodiester bonds between teichoic acid monomers. Outside the cell wall, many Gram-positive bacteria have an S-layer of "tiled" proteins. The S-layer assists attachment and biofilm formation. Outside the S-layer, there is often a capsule of ...
There is no connection between the shape of a bacterium and its color upon Gram staining; there are both gram-positive rods and gram-negative rods. MacConkey agar can be used to distinguish among gram-negative bacilli such as E. coli and salmonella. [23]
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.
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.
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.