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The Gram stain is a general stain used extensively in microbiology for the preliminary differentiation of microbiological organisms. The Gram stain is one of the simplest, least expensive, and most useful of the rapid methods used to identify and classify bacteria.
The Gram stain is used to provide preliminary information concerning the type of organisms present directly from clinical specimens or from growth on culture plates. This stain is used to identify the presence of microorganisms in normally sterile body fluids (cerebrospinal fluid, synovial fluid, pleural fluid, peritoneal fluid). It is also used to screen sputum specimens to establish acceptability for bacterial culture (<25 squamous epithelial cells per field is considered an acceptable specimen for culture) and may reveal the causative organism in bacterial pneumonia.
Identifying microorganisms in normally sterile body fluids
Screening sputum specimens for acceptability for bacterial culture
Guiding initial antimicrobial therapy
During the staining process, the crystal violet and iodine form a complex within the heat fixed cell. In gram-negative organisms, this complex is readily washed out by the acetone-alcohol. They appear red because they retain only the safranin dye (counterstain). Gram-positive organisms retain the crystal violet-iodine complex after decolorization and remain purple.
Overdecolorization may result in the loss of the crystal violet iodine complex from gram-positive organisms and result in a misinterpretation.
No organisms seen or descriptive report of observations.
Atlas RM, Snyder JW: Reagents, Stains, and Media: Bacteriology. In Manual of Clinical Microbiology. Vol 1. 10th edition, Edited by J Versalovic. Washington DC, American Society for Microbiology, 2011, pp 272-307