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The Gram stain is the most common bacteriologic stain used in the clinical laboratory. It consists of the application of a crystal violet solution to a heat-fixed bacterial smear on a glass slide for 60 seconds. The slide is then rinsed gently in distilled water, and an iodine solution is added for 30 seconds. The slide is again rinsed, and a solution of primarily ethanol is rinsed over the slide for 10–15 seconds. The slide is again rinsed, and a solution of safranin is added for one additional minute. After the slide is rinsed a final time and dried, it is then observed with a microscope at 1000× magnification.
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