Alcohol in Gram staining removes stains from Gram-negative cells, and Gram-positive cells remain purple after the procedure.
Alcohol serves as a decolorizer in the Gram stain process. It removes the crystal violet/iodine complex from Gram-negative cell walls, distinguishing them from Gram-positive cells.
After the Gram stain, Gram-positive cells remain purple, while Gram-negative cells lose the purple color and appear transparent. The addition of a counterstain like safranin gives Gram-negative cells a pink color for visualization.
Timing during the decolorizing step is crucial to prevent misclassification of bacterial cells.
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