What is the Decolorizer used in the Endospore stain?
What is the Decolorizer used in the Endospore stain?
It takes a long time for the spores to stain due to their density, so time acts as the mordant when performing this differential stain; the slide with the bacterium should be soaked in malachite green for at least 30 minutes and then rinsed off with water which acts as the decolorizer.
What are some of the applications of Endospore staining?
Applications of the Endospore stain
- For detection of Firmicute groups of bacteria i.e Clostridium spp and Bacillus spp.
- For identification of endospore producing bacteria in samples.
- for differentiation of spore-producing bacterial from vegetative forms of bacteria.
What genuses form endospores?
Endospores are dormant alternate life forms produced by the genus Bacillus, the genus Clostridium, and a number other genera of bacteria, including Desulfotomaculum, Sporosarcina, Sporolactobacillus, Oscillospira, and Thermoactinomyces.
Why is there no Decolorizer in this spore stain?
Endospores are not for reproduction: One spore forms inside of the vegetative cell. When the spore germinates, one vegetative cell will be produced. That is why there does not need to be a decolorizer in this stain: it is based on the binding of the malachite green and the permeability of the spore vs. cell wall. .
How long can endospores survive?
Endospores enable bacteria to lie dormant for extended periods, even centuries. There are many reports of spores remaining viable over 10,000 years, and revival of spores millions of years old has been claimed.
What is the principle of spore stain?
Principle: Endospore staining is a differential staining technique where the spore is stained in a manner so that it can be distinguished from the vegetative part of the cell. Spores are structures remarkably resistant to heat, radiation, chemicals and other agents that are typically lethal to the organism.
What is the function of an endospore?
It allows the bacterium to produce a dormant and highly resistant cell to preserve the cell’s genetic material in times of extreme stress. Endospores can survive environmental assaults that would normally kill the bacterium.
What is the main function of endospores?
What bacteria Cannot form endospores?
Listeria monocytogenes is a Gram-positive rod-shaped bacterium related to Bacillus and Clostridium, but it does not form endospores.
How do you identify an endospore?
Lastly, in endospore staining, Vegetative cells will be stained red by the safranin counter stain. If endospores are present in the sample, these will retain the malachite green stain, and appear bluish-green in color.
Which is the best decolorizer for endospore staining?
( Note: In Gram Staining and AFB Staining we use Alcohol or Acid Acohol or Acid as a decolorizer but in spore staining water is sufficient ( to be used as decolorizer) because: malachite green dye is water-soluble and does not adhere well to the cell wall vegetative cells have been disrupted by heat,
How is malachite green used as an endospore stain?
The primary stain in the endospore stain procedure, malachite green, is driven into the cells with heat. Since malachite green is water-soluble and does not adhere well to the cell, and since the vegetative cells have been disrupted by heat, the malachite green rinses easily from the vegetative cells, allowing them to readily take up…
How are spores formed in an endospore stain?
Endospore Stain . The endospore stain is a differential stain used to visualize bacterial endospores. Endospores are formed by a few genera of bacteria, such as Bacillus . By forming spores, bacteria can survive in hostile conditions. Spores are resistant to heat, dessication, chemicals, and radiation.
Can A Schaeffer Fulton stain be used on an endospore?
Endospores cannot be stained by normal staining procedures because their walls are practically impermeable to all chemicals. The Schaeffer- Fulton endospore stain uses heat to drive the primary stain (malachite green) into the endospore. After cooling, the slide is decolorized with water and counterstained with safranin.