Gram staining consists of three processes...

Gram-negative staining Created What gram staining? Gram staining common method used to distinguish two major groups of bacteria based on their different components of the cell wall. Gram procedure distinguishes between gram-positive and gram-negative group on coloring the cells of red or purple. Gram-positive bacteria stain purple due to a thick layer of peptidoglycan in their cell wall that retains crystal violet stained these cells. In addition, gram-negative bacteria stain red, due to a thin peptidoglycan wall that does not keep crystal violet during the bleaching process. How grams of Painting? Gram staining consists of three processes: painting water-soluble dye called crystal violet decolorization and counterstaining, usually with safanin. Because of variations in layer thickness of peptidoglycan in the cell membrane between gram-positive and gram-negative bacteria, gram-positive bacteria (more than a thick layer of peptidoglycan) retain crystal violet stain during the bleaching process, while Gram-negative bacteria lose the crystal violet stain and instead painted safranina final process of painting. The process includes three stages:

cells stained with the dye crystal violet. Further, the solution g of iodine (iodine and potassium iodide) is added to form a complex between crystal violet and iodine. This complex is a large molecule than the original crystal violet stain and iodine and insoluble in water. Decolorizer such as ethanol or acetone is added to the sample, which dehydrates peptidoglycan layer, reduction and strengthening it. Large crystal violet iodine complex can not penetrate this increased peptidoglycan layer, and thus trapped in a cage in gram-positive bacteria. On the other hand, the outer membrane of gram-negative bacteria is reduced and the thinner layer of peptidoglycan Gram-negative cells can not retain crystal violet iodine complex and the color is lost. Counterstain, for example, poorly soluble in water safranina, added to the sample zabarvlyuyuchy it in red. With safranina easier than crystal violet, it does not violate the purple color in gram-positive cells. However, discolored Gram-negative cells stained red. As staining protocol and issues: Reagents: Decolorizer (eg, ethanol)

Make slide cell samples should be painted. Heat fix the slide master carefully passing the slides from falling or a small piece of the sample on it through the burner three times. Add the primary stain (crystal violet) for the sample / slide and incubate for 1 minute. Rinse the slide with a gentle stream of water for no more than 5 seconds to remove unbound crystal violet. Add grams of iodine for 1 minute, a hydrate, or agent, which fixes the crystal violet to bacterial cell wall. Rinse the sample / slide with acetone or alcohol within ~ 3 seconds and strattera online rinse with running water. Alcohol illuminate samples, where Gram-negative, removal of crystal violet. However, if

alcohol remains in the sample is too long, it can also illuminate the gram-positive cells. Add the secondary spot safranina on slide and incubate for 1 minute. Rinse spray water for no longer than 5 seconds. If gram-positive bacteria, it will keep the main color (crystal violet), and do not take secondary stain (safranina), making it look purple / purple under the microscope. If gram-negative bacteria, he loses the primary colors and make secondary spots, causing appear red when viewed under a microscope. Links literature and educational activities.