![]() To prepare a smear from a broth culture, instead of carrying out steps 2 and 3, aseptically remove several loopfuls of culture and spread over the circled area.Įxpect the smear to be far less dense than smears prepared from colonies on agar.The hottest part of the flame is just above the tip of the inner cone. ![]() To obtain a sufficiently hot flame, adjust the air-gas mixture so that the gas burns with a pale blue flame (inner cone) with a nearly colorless large outer cone.Make sure that the liquid is completely dry before passing the slide through the flame.Heating the slide briefly partially melts the cell walls, causing the cells to adhere to the glass surface.Let the slide cool for 30 sec or so before starting the staining procedure.Each smear should pass through the hottest part of the flame (see the third note below) and each pass through the flame should take about a second. Hold one end of the slide horizontally with a clothes pin and pass it through a flame three times with the smears up.Allow the drop to air dry completely (usually a couple of minutes for a single loopful).Try to disperse the culture material completely, so that there are no visible chunks of material, and spread the liquid into a thin layer within the circle. ![]()
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