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Use of the Gram stain in microbiology
Authors:TJ Beveridge
Institution:  a Department of Microbiology, College of Biological Science, University of Guelph, Guelph, Ontario, Canada.
Abstract:The Gram stain differentiates bacteria into two fundamental varieties of cells. Bacteria that retain the initial crystal violet stain (purple) are said to be 'Gram-positive,' whereas those that are decolorized and stain red with carbol fuchsin (or safranin) are said to be 'Gram-negative.' This staining response is based on the chemical and structural makeup of the cell walls of both varieties of bacteria. Gram-positives have a thick, relatively impermeable wall that resists decolorization and is composed of peptidoglycan and secondary polymers. Gram-negatives have a thin peptidoglycan layer plus an overlying lipid-protein bilayer known as the outer membrane, which can be disrupted by decolorization. Some bacteria have walls of intermediate structure and, although they are officially classified as Gram-positives because of their linage, they stain in a variable manner. One prokaryote domain, the Archaea, have such variability of wall structure that the Gram stain is not a useful differentiating tool.
Keywords:Carbol Fuchsin  Crystal Violet  Gram-NEGATIVE Bacteria  Gram-POSITIVE Bacteria  Gram Stain  Gram-VARIABLE  Iodine
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