thank you in this video you will learn how to perform gram staining this technique utilizes a four-step staining procedure with two different dies and is one of the first tests used when trying to identify unknown species of bacteria using a wax pencil label a microscope slide with your initials then draw a circle about one centimeter in diameter on the slide using a metal inoculation Loop sterilize it by flaming using aseptic technique add a drop of sterile water so that a film appears across the loop transfer the water into the circle drawn by the wax pencil Flame the metal loop again to sterilize foreign it to cool use the metal Loop to transfer a colony of bacteria growing on an auger plate make sure to lightly touch the bacterial colony to avoid transferring too many bacteria when finished Flame the loop to sterilize let the liquid completely evaporate leaving behind a dry spot on the slide this will take at least 10 minutes once the liquid has dried completely heat fix the bacteria to the slide by quickly passing the slide through the flame three times the flame should touch the slide for about one second during each pass ensure that all Bunsen burners are turned off before obtaining the set of gram stains the decolorizer contains a high percentage of alcohol and is very flammable add a few drops of Crystal Violet stain to the bacteria until the wax circle is filled with stain let this stand for one minute rinse the slide by dipping it in a beaker of water two to three times until most of the stain has been washed off next add a few drops of grams iodine to cover the bacteria and then let this stand for one minute rinse the slide in a beaker of water until most of the iodine is gone a second Beaker of water can be used to ensure most of the iodine has been rinsed off now add a few drops of decolorizer let the decolorizer flow over the bacteria at a 45 degree angle until the flow is clear then finish by rinsing the slide in a beaker of water foreign by adding safranin stain to cover the bacteria and let it stand for one minute after one minute completely rinse the slide in water allow the slide to dry completely once the slide is completely dry observe the sample under a microscope