Why 95% alcohol is used in Gram staining?

Remel Gram Decolorizer (95% Ethyl Alcohol) is a reagent recommended for use in qualitative procedures to differentiate gram-negative from gram- positive organisms. The primary stain, crystal violet, is a basic dye which rapidly permeates the cell wall of all bacteria, staining the protoplast purple.Click to see full answer. In this way, why alcohol is used in Gram staining?A decolorizer such as ethyl alcohol or acetone is added to the sample, which dehydrates the peptidoglycan layer, shrinking and tightening it. The large crystal violet-iodine complex is not able to penetrate this tightened peptidoglycan layer, and is thus trapped in the cell in Gram positive bacteria.Additionally, why Iodine is used in Gram staining? Gram staining is based on the ability of bacteria cell wall to retaining the crystal violet dye during solvent treatment. Bacteria cell walls are stained by the crystal violet. Iodine is subsequently added as a mordant to form the crystal violet-iodine complex so that the dye cannot be removed easily. Also know, why is the decolorization important in Gram staining? The Gram stain is the most important staining procedure in microbiology. This layer makes up 60-90% of the gram positive cell wall. Decolorizing the cell causes this thick cell wall to dehydrate and shrink, which closes the pores in the cell wall and prevents the stain from exiting the cell.What is the most important reagent in Gram stain method?The primary stain of the Gram’s method is crystal violet. Crystal violet is sometimes substituted with methylene blue, which is equally effective. The microorganisms that retain the crystal violet-iodine complex appear purple brown under microscopic examination.