Differential Staining Essay

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Differential Staining Brenda Blanchette, RN, CCTC Clinical Microbiology Laboratory Differential Staining There are several staining methods used with bacteria and are generally classified as simple, nonspecific, or differential (specific). Gram’s stain techniques are used to differentiate between types of bacteria as well as to differentiate between Gram-positive and Gram-negative bacteria. Gram’s stains are one of the most commonly used differential stains and are the focus of this experiment (Labpaq microbiology). Bacteria that stain purple are Gram-positive and bacteria that stain pink are Gram-negative. They stain differently due to differences in the structure of their cell walls. Gram-positive bacteria stain purple because they retain the crystal violet dye after the alcohol and acetone wash of the Gram stain process. Gram-positive bacteria have very thick cell walls of which about 60%- 90% of the wall is peptidoglycan. They also have teichoic acids interwoven in the wall (Betsy, T., & Keogh, J. 2005). Gram-negative bacteria stain pink because after the wash of alcohol and acetone they do not retain the crystal violet dye leaving them decolorized. A counterstain of safranin is added which stains the bacteria pink. Gram-negative bacteria have only a few layers of peptidoglycan that are surrounded by an outer membrane of phospholipids, lipopolysaccharide and lipoprotein. There is a space between the peptidoglycan and the secondary cell membrane called the periplasmic space (Betsy, T., & Keogh, J. 2005). Initially Gram staining begins with the basic crystal violet dye and both Gram-positive and Gram-negative appear purple. Then the bacteria are treated with Gram’s iodine, a mordant used in gram staining to intensify a stain (Betsy, T., & Keogh, J. 2005). The mordant helps retain the stain by forming an insoluble
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