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In microscopy, negative staining is an established method, often used in diagnostic microscopy, for contrasting a thin specimen with an optically opaque fluid. In this technique, the background is stained, leaving the actual specimen untouched, and thus visible. This contrasts with positive staining, in which the actual specimen is stained.
A simple staining method for bacteria that is usually successful, even when the positive staining methods fail, is to use a negative stain. This can be achieved by smearing the sample onto the slide and then applying nigrosin (a black synthetic dye) or India ink (an aqueous suspension of carbon particles).
Gram-negative bacteria are bacteria that, unlike gram-positive bacteria, do not retain the crystal violet stain used in the Gram staining method of bacterial differentiation. [1] Their defining characteristic is that their cell envelope consists of a thin peptidoglycan cell wall sandwiched between an inner ( cytoplasmic ) membrane and an outer ...
Gram stain (Gram staining or Gram's method), is a method of staining used to classify bacterial species into two large groups: gram-positive bacteria and gram-negative bacteria. It may also be used to diagnose a fungal infection. [1] The name comes from the Danish bacteriologist Hans Christian Gram, who developed the technique in 1884. [2]
Gram-negative bacteria will stain a pink color due to the thin layer of peptidoglycan. If a bacteria stains purple, due to the thick layer of peptidoglycan, the bacteria is a gram-positive bacteria. [4] In clinical microbiology numerous other staining techniques for particular organisms are used (acid fast bacterial stain for mycobacteria).
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.
Other bacteria are commonly identified with a microscope by staining them with Gram stain. However, the mycolic acid in the cell wall of M. tuberculosis does not absorb the stain. Instead, acid-fast stains such as Ziehl–Neelsen stain, or fluorescent stains such as auramine are used. [4]
In Gram-negative bacteria an outer membrane is also included. [1] This envelope is not present in the Mollicutes where the cell wall is absent. Bacterial cell envelopes fall into two major categories: a Gram-positive type which stains purple during Gram staining and a Gram-negative type which
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