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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.
Gram staining. Gram staining or Gram stain, also called Gram's method, is a method of staining used to differentiate bacterial species into two large groups (gram-positive and gram-negative). The name comes from the Danish bacteriologist Hans Christian Gram, who developed the technique.