![]() ![]() Prepare smears of organisms to be tested for the presence of endospores on a clean microscope slide and air dry it.Mature, free endospores should not be associated with vegetative bacteria and should be seen as green ellipses.the vegetative cells containing endospores should stain pink, while the spores should be seen as green ellipses.the vegetative cells should appear pink/red (i.e., the color of counterstain),.When visualized under microscopy, the cells should have three characteristics: When counter-stained with safranin, the vegetative cells take the color of safranin and appear red or pink, in contrast to the endospores that appear green. Once the endospore has absorbed the stain, it is resistant to decolorization, but the vegetative cells are easily decolorized with water (leaving the vegetative cells colorless). In this technique, heating acts as a mordant. When a heat-fixed smear is flooded with aqueous malachite green solution (the primary stain) and steamed, the heat assists the stain in penetrating through the spore. The method utilizes malachite green as the primary stain and safranin as a counterstain. Schaeffer and MacDonald Fulton in the 1930s. The technique was first described by Alice B. It is the most widely used technique for endospore staining. They appear as large refractile oval or spherical bodies within the mother cell. Endospores can also be demonstrated in unstained wet films under a phase-contrast microscope. Spores can generally be recognized on Gram’s stains (endospores do not stain and appear as refractile, nonstaining bodies). ![]() There are different methods for endospore staining. Other techniques of endospore staining Methods for endospore staining.Principle of Dorner’s method for staining endospores. ![]()
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