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Gram Staining: A. Add crystal violet stain over the fixed culture. Let stand for 10 to 60 seconds; for thinly prepared slides, it is usually acceptable to pour the stain on and off immediately. Pour off the stain and gently rinse the excess stain with a stream of water from a faucet or a plastic water bottle. Note that the objective of this step is to wash off the stain, not the fixed culture. B. Add the iodine solution on the smear, enough to cover the fixed culture. Let stand for 10 to 60 seconds.
In the staining process, those that decolorize easily are gram negative, and the one that retain the primary stain are gram positive.   The acid fast staining is a techique that stains only the gram positive and only the genuses of mycobacterium, nocardia, and actinomycetes.ram staining   Gram Staining is a way to separate one large groups of bacteria into two. Crystal violet is used to dye the cells, those that retain the color are grouped â¦ as Gram-positive, and those that do not retain the color are grouped as Gram-negative.