Answer Posted / r.rajani
Gram staining is based on the ability of bacteria cell wall
to retaining the crystal violet dye during solvent
treatment. The cell walls for Gram-positive microorganisms
have a higher peptidoglycan and lower lipid content than
gram-negative bacteria. 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. This step is commonly
referred to as fixing the dye. However, subsequent treatment
with a decolorizer, which is a mixed solvent of ethanol and
acetone, dissolves the lipid layer from the gram-negative
cells. The removal of the lipid layer enhances the leaching
of the primary stain from the cells into the surrounding
solvent. In contrast, the solvent dehydrates the thicker
Gram-positive cell walls, closing the pores as the cell wall
shrinks during dehydration.
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