Heat fix the smear by running the slide through the flame about 20 times, and leave malachite green on for 20 minutes during the stain process. Notice where the spore is in the cell, if there is oneterminal, central, subterminal.
This may be helpful information in the identification of the unknown. Objectives Learn to perform the spore stain. Identify spores on a bacterial smear.
Put a beaker of water on the hot plate and boil until steam is coming up from the water. Then turn the hot plate down so that the water is barely boiling. Place the wire stain rack over the beaker which now has steam coming up from the boiled water. Cut a small piece of paper towel and place it on top of the smear on the slide. The towel will keep the dye from evaporating too quickly, thereby giving more contact time between the dye and the bacterial walls.
Flood the smear with the primary dye, malachite green, and leave for 5 minutes. Keep the paper towel moist with the malachite green. DO NOT let the dye dry on the towel. Remove and discard the small paper towel piece. Endospores are formed within a vegetative bacterial cell when the environmental conditions no longer support cell growth. As the vegetative cell dies, the endospore is released into the environment where it can survive indefinitely in the presence of many environmental stresses, such as desiccation, extremes in temperature, radiation, and lack of nutrients.
When more favorable conditions arise the endospore germinates, again forming a viable vegetative cell. The presence of endospores in a bacterial culture can be detected by staining with malachite green. Because the endospore coat is so tough, steam is used to enable dye penetration. After washing, only the endospores will retain the primary stain Malachite green.
Safranin is then used as a counterstain for vegetative cells. The endospore stain is a differential stain because it differentiates spore-formers from non spore -formers. Note: Formation of an endospore. The spore stains green and the vegetative cells stain red to orange. Last updated on June 21st, Bacterial spores are highly resistant, dormant structures i.
When vegetative cells of certain bacteria such as Bacillus spp and […]. News Ticker. Contents 1 Methods for endospore staining 1. Gram Positive Rods. About Acharya Tankeshwar Articles. Hello, thank you for visiting my blog. I am Tankeshwar Acharya. Blogging is my passion. I am working as an Asst.
Anaerobic Bacteriology. Acharya Tankeshwar Bacteriology , Microbiology for Beginners 7.
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