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It acts as the mordant to soften the mycolic acid so that the stain can penetrate the cell.

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Waldo Mayer

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2y ago

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What is the purpose of staining the smear in malachite green during spore staining?

the purpose of boiling of smear in malachite green is to forces a stain to penetrate the endospore wall, it is necessary to heat the slide and the stain to prod the wall to allow the stain to enter.


Why is it you should heat-fixing the air-dried before staining?

It is used to fix because to make the cell inactive or immoblie, but the main purpose is to fix the smear so that when we put stain and then flush it out with water ( or some time with alcohol) the smear should not wash out with dye.


What is the purpose of prolonged application of heat on the smear?

by application of heat and a mordant, the cell can be stained. the purpose of heating is to soften the waxy material of the cell wall and allow the stain enter the cell. rock on University of Luzon!


During the performance of the simple staining procedure you failed to heat fix your EColi smear preparation Upon microscopic examination how would you expect this slide to differ from correct slide?

Without heat fixing, the bacteria on the slide will not adhere properly, leading to poor attachment to the slide during staining. This may result in uneven staining, leading to difficulty in visualizing the bacterial cells clearly under the microscope. Proper heat fixing ensures that the bacteria are securely attached to the slide, allowing for better staining and clearer observation under the microscope.


What would happen if no heat fixing were done?

If no heat fixing was done to a slide with a specimen on it, it would be rinsed off with the gram staining procedure. Heat fixing the specimen does kill specimen but it also locks it in place.


Which staining procedure uses heat to drive the stain in?

The heat-based staining procedure is called heat fixation. In this process, heat is used to adhere the specimen to the slide, allowing it to withstand the subsequent staining and washing steps without washing away.


Why heat slide in aceto-orcein staining?

Heating during aceto-orcein staining enhances the permeability of the cellular membranes, allowing the dye to penetrate more effectively. This process helps to intensify the staining of nucleic acids, making chromatin structures more visible under a microscope. Additionally, heat can facilitate the fixation of the dye to the cellular components, improving the overall quality of the staining.


What is the function of a Bunsen burner in microbiology?

In microbiology, a Bunsen burner is used to sterilize tools and heat-fix bacterial smears onto slides for staining purposes. The flame produced by the Bunsen burner provides a sterile environment to prevent contamination during microbiological procedures.


When is heat fixing appropriate to a negative stain on a bacterial sample?

Heat fixing is generally not appropriate for negative staining because it can alter the shape and size of the bacterial cells, leading to inaccurate results. Negative staining relies on the use of acidic dyes that do not penetrate the cells, allowing for clear visualization of the cell's morphology and size against a contrasting background. Heat fixing can cause cells to shrink or distort, which defeats the purpose of using a negative stain. Therefore, it is best to avoid heat fixing when preparing samples for negative staining.


What are the advantage and disadvantage of heat fixing?

Advantages: It helps adhere bacterial cells to the slide, preventing them from washing away during staining. Also, it kills the bacteria, making them safe to handle and study under the microscope. Disadvantages: Heat fixing can distort the morphology of the bacterial cells, affecting the accuracy of the staining results. Overheating can also cause cell lysis, leading to inaccurate interpretation of the specimen.


What is a primary stain?

If your talking about Acid Fast staining (aka Ziehl-Neelsen staining), after the addition of the primary stain (carbol fuchsin) heat is applied in order to facilitate proper staining. Bacteria such as Mycobacterium contain large amounts of lipid substances within their cell wall called mycolic acids. These acids resist staining by ordinary methods such as gram staining (where heat is not applied after primary staining). On application of heat, the stain carbol fuschin penetrates the cell wall and stains the cells pink. Following the secondary staining (methylene blue) the acid fast positive cells appear pink while others are stained blue. Endospore staining is yet another staining technique where heat is applied after primary staining (malachite green). In this case the spores are impermeable to stains, hence heating favours proper permeation of stain. Endospores appear green while vegetative cells appear red (secondary stain saffranine). Not all staining procedures involve applying heat after primary staining. However, heat is applied before even beginning the staining procedure. This is called heat fixing, where the cells are fixed so that they are not washed away during the subsequent washing steps.


Function of heat-fix in gram staining?

Heat-fixing in gram staining serves to adhere bacterial cells to the slide, making them more resistant to washing off during the staining process. It also helps to kill the bacteria, allowing them to take up the crystal violet stain more effectively. Additionally, heat-fixing can alter the permeability of the bacterial cell wall, aiding in the retention of the stain through subsequent decolorization steps.