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because giemsa stain is a mixture of methyl acetate Eosin and azure b. it doesnot contain any fixative that is why we use methanol to fix smear during giemsa stain
other stain like lieshman contain acetyl free methyl alcohol as a fixative so it does not need to fix slide stain with lieshman stain.
Methanol is used to fix the smear during Giemsa staining because it helps to preserve the cellular structure by denaturing proteins and preventing their degradation. It also permeabilizes the cells, allowing for better penetration of the dye and increased staining intensity. Additionally, methanol evaporates quickly, which helps to reduce the staining time and allows for quicker results.
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Give the reason for passing bacterial smear through the flame before staining?
Passing the bacterial smear through the flame before staining is done to heat-fix the bacteria onto the slide, making them adhere firmly and preventing them from washing off during the staining process. Heat fixing also kills the bacteria, which helps in the preservation of their cellular structures for visualization under the microscope.
What is the function of methanol in the wright's stain solution?
Methanol is used in Wright's stain solution as a solvent to help dissolve the dyes and facilitate their penetration into cells, tissues, or other biological samples for staining purposes. It also helps to fix the stain onto the sample by enhancing the adhesion of the dye to the cellular components.
What happens if you don't heat fix a smear prior to staining of a bacterial slide?
If a bacterial smear is not heat fixed prior to staining, the bacteria may not adhere well to the slide and can wash away during the staining process. Heat fixing helps to kill the bacteria, firmly attach them to the slide, and improve the uptake of stain, resulting in better staining results. Without heat fixing, the bacteria may not stain properly or may not be visible at all under the microscope.
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.
Why is it you should heat-fixing the air-dried before staining?
Heat-fixing the air-dried smear before staining helps the cells adhere to the slide better, preventing loss of specimen during staining procedures. It also helps in denaturing and killing the bacteria or other microorganisms, making them more visible and easier to stain.
Why acetone free methanol in lieshmans stain?
Acetone-free methanol is used in Lillie's modified Lieberman's iron hematoxylin (Lieshman stain) because acetone can cause precipitation of the hematoxylin pigment and thus affect staining quality. Methanol is often preferred over acetone for its effective solvent properties and compatibility with the staining process.
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.
Give the reason for passing bacterial smear through the flame before staining?
Passing the bacterial smear through the flame before staining is done to heat-fix the bacteria onto the slide, making them adhere firmly and preventing them from washing off during the staining process. Heat fixing also kills the bacteria, which helps in the preservation of their cellular structures for visualization under the microscope.
What is the function of methanol in the wright's stain solution?
Methanol is used in Wright's stain solution as a solvent to help dissolve the dyes and facilitate their penetration into cells, tissues, or other biological samples for staining purposes. It also helps to fix the stain onto the sample by enhancing the adhesion of the dye to the cellular components.
What happens if you don't heat fix a smear prior to staining of a bacterial slide?
If a bacterial smear is not heat fixed prior to staining, the bacteria may not adhere well to the slide and can wash away during the staining process. Heat fixing helps to kill the bacteria, firmly attach them to the slide, and improve the uptake of stain, resulting in better staining results. Without heat fixing, the bacteria may not stain properly or may not be visible at all under the microscope.
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.
Why is it you should heat-fixing the air-dried before staining?
Heat-fixing the air-dried smear before staining helps the cells adhere to the slide better, preventing loss of specimen during staining procedures. It also helps in denaturing and killing the bacteria or other microorganisms, making them more visible and easier to stain.
Why do you air-dry a smear?
Air-drying a smear helps to fix the cells onto the slide, preventing any loss or distortion during further processing steps like staining or examination under a microscope. It also helps to evaporate any excess water, improving the visualization of the cells.
What is the effect of heating the smear flooded with a carbol fuchsin stain?
Heating the smear flooded with carbol fuchsin stain helps in the penetration of the stain into the bacterial cell wall by softening the cell wall and making it more permeable. This process is important for the retention of the stain during the subsequent decolorization step in the staining process.
What is the purpose of prolonged application of heat on the smear?
Prolonged application of heat on a smear during staining helps to improve the uptake of the stain by bacterial cells and enhances the contrast between different cell types. The heat can also help to fix the cells to the slide, preventing them from being washed away during subsequent steps in the staining process.
What is gram variability?
Gram variability refers to a characteristic of certain bacteria that can exhibit variability in their response to Gram staining, appearing as a mix of both Gram-positive and Gram-negative characteristics. This variability can make the identification of these bacteria challenging because their staining characteristics may not be consistent.
In gram staining why will very thick smears give inaccurate results?
Very thick smears in Gram staining may lead to inaccurate results because the dye may not penetrate all layers of the smear evenly. This can result in difficulties distinguishing between Gram-positive and Gram-negative bacteria, leading to ambiguous or incorrect results. It is important to prepare thin and evenly spread smears to ensure accurate staining and interpretation.
