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What general type of stain is used to separate types of bacteria based on their cellular structures?

Question

What general type of stain is used to separate types of bacteria based on their cellular structures?

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Solution

The general type of stain used to separate types of bacteria based on their cellular structures is called a differential stain. The most common differential stains are the Gram stain and the Acid-fast stain.

Here are the steps for the Gram stain:

  1. A heat-fixed smear of bacterial culture is prepared on a microscope slide.
  2. Crystal violet (the primary stain) is added to the smear. All cells stain purple at this point.
  3. Iodine (the mordant) is added. This forms a complex between the crystal violet and the iodine.
  4. Alcohol or acetone (the decolorizer) is added. This dehydrates the peptidoglycan layer, shrinking and tightening it. Gram-positive bacteria retain the crystal violet stain, and gram-negative bacteria do not.
  5. Safranin (the counterstain) is added. This stains decolorized gram-negative bacteria pink or red.

The Acid-fast stain is used for a few types of bacteria that do not take up the Gram stain (like Mycobacterium tuberculosis). The steps are similar, but the primary stain is carbol fuchsin, the decolorizer is acid-alcohol, and the counterstain is methylene blue. Acid-fast bacteria retain the carbol fuchsin stain even in the presence of acid-alcohol and are pink, while non-acid-fast bacteria are blue.

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