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Spores are very hard and dense, dye is not readily absorbed into the endospore. However, one method of staining is the Schaeffer and Fulton method. The stain is malachite green and the proper method entails preparing a heat fixed smear which is covered by a piece of blotting paper, and flooded with the dye. Wait 15 mins then remove blotting paper and wash. Counterstain with 0.5% Safrinin.
Spores appear green.
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Of what use is the endospore stain?
The use of endospore stain is to see specialized cell structures. It can tell if some bacterium cells contain higher resistant spores within vegetative cells.
How does culture age affect the results of a sproe stain?
Spores are formed when cells are under unfavourable conditions, as for the bacteria they are means of survival. So the older the culture the higher the cell number in that culture, which means less nutrients for the cells. Under this conditions cells will start spore production. Depending on the "age" of the culture you can get a mixture of vegetative cells with spores inside and spores that are already released or mostly spores with rare vegetative cells, which means the cells are dead.
What prevents the cell from appearing green in the finished endospore stain?
Vegetative cells readily decolorize because they are permeable.
Assume you stain Bacillus by applying malachite green with heat and then counterstaining with safranin Through the microscope the green structures are?
The malachite green penetrates the endospores, the counterstain of safranin will stain the vegetative cells. Endospores - greenish-blue Vegetative Cells - Red/Pink
Can you see spores in a gram stain preparation?
yes, spore will not stain , so it will shown as a empty space ( bright refrigent )inside the cells . the rest of the cells will be purple colored.
Why is it necessary to perform a spore stain when you can see the presence of spores in simple stained cells?
spore is a dormant structure and necessary to stain because in this we use malachite green which will retain in spore
What are the properties that differentiate the spore from the vegetative cell?
In the spore stain malachite green is used as a primary stain. This is driven into the cell by heat because of the impermeability of the spore. The stain is allowed to sit for 30 mins to make sure it gets in to the endospores.The stain is then washed and counterstained with safranin red. The endospores retain the green colour from malachite green and of course appear green under the microscope. Whereas the vegetative cells will appear red.
Why are spores so difficult to stain?
These bacteria that have flagella have to be handled more carefully than bacteria without flagella because the flagella could brake off the bacteria. Remember that some bacterial flagella are very small and delicate.
How would an endospore stain of mycobacterium appear?
Acid fast Mycobacterium have a waxy molecule in their cell wall that will take up and retain the malachite green stain when subjected to the endospore staining process. The uniformly green appearance of endospore stained acid fast cells doesn't mean they produce endospores. These are vegetative cells that have taken up color from the heat driving malachite green into their waxy cell wall.
Is endospore staining differencial staining?
Endospore staining is a differential stain used to detect the presence and location of spores in bacterial cells.
What kind of stain are used to stain plant and animal cells?
Methylene blue stain is used to stain plant and animal cells.
Why do all cells stain purple in the flagella stain but not in the gram stain?
The gram stain uses a decolorizing product so it is possible to differentiate between the gram and the gram cells. Gram positive cells stain purple in color.
