why prolong heat fixing is important in spore stainin
Answers
Answer:
In basic Laboratories, the simplest endospore staining technique is the Schaeffer-Fulton technique because of its easy and it is rapid to identify the bacteria. It applies the use of Malachite green dye (alkaline solution with a pH of 11-11.2 and its a water-soluble dye) along with the use of steamed-heat which softens the endospore covering allowing penetration of the dye into the spore. The malachite green dye binds to the spore mildly and if washed with water, without fixing, it easily washes away, and that’s why the application of steamed heat is important to allow the dye to penetrate the endospore. Water is used as a decolorizing agent, to was away from the malachite dye from vegetative forms. Lastly, the use of a counterstain, Safranin reagent, also knows as the secondary stain, is to stain the vegetative forms of the underdeveloped Furmigates vegetative forms after the malachite dye has been washed away by the decolorizing agent (Water).
Malachite Green dye and Safranin work well in bacteria because of the alkaline nature of the Malachite Green reagents which are charged positively while the cytoplasm of the bacterial cell is basophilic hence there is an attraction between the malachite green dye with the bacterial cell, making it easier to absorb the dye. Visualization of the cells under the microscope will show the appearance of pink-red stain for the vegetative cell forms, which take up the counterstain while the endospores will appear as green dotted particles (ellipses), having taken up the Malachite green dye.
Answer:
Apply heat to assist stain penetration. the heat will force the stain into the spores and vegetative cells and both will become green. by rinsing the slids, the vegetative cells will lose their color but the spores will remain green
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