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.
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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.
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.
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.
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.
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.
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.