Question

In order to prepare an optimal heat-fixed bacterial smear for staining, one must be aware of a couple of factors that may lead to poor results; briefly explain these. Why is it necessary to heat fix a smear for simple staining?

Answer 1

Answer:

The answer is below

Explanation:

Some of the factors that may lead to poor results in an optimal heat-fixed bacterial smear for staining are the following:

1.  Thickness of the smear: In a situation whereby the smear is too thick, it would be impossible to perform adequate decolorization.

2. Quantity of Decolorizer: if the quantity of the decolorizer or reagent is concentrated, the quality of the stain will be affected

3.  Age of the culture: Older cultures have multiple ruptured and dead cells, which result in staining Gram-negative.

4. Duration of Decolorization:  Overheated smear during heat fixing, often leads to the rupture of cell walls.

The reason it is necessary to heat fix a smear for simple staining is that:

Heat fixing affects bacterial enzymes original state, which hinders their cell digestion parts, thereby leading to the breaking of the cell. The heat also improves the steadfastness of bacterial cells to the slide.

Answer 2

It is essential to heat and fix a smear for simple staining as it helps the bacteria to get attached with the slide. However, one must also be aware of the factors, which may lead to poor results.

What is heat fixation?

It is a laboratory method used commonly in histology, cell biology, and pathology in which a slide with a specimen like bacteria is passed via the source of heat for many times.

It is an essential step in the procedure of staining as the heat fixation denatures the bacterial enzymes that can autolyze the cells of bacteria, therefore, to inhibit this there is a need to heat fix the smear. The heat fixation also enhance the attachment between the bacteria and the glass of slide so by which one can witness the smear clearly under the microscope.

Apart from these, there are some factors, which can result in poor results. Like when mixing bacteria with water, if one overmix, it can lead to inaccurate cell patterns. At the time of heat fixing, if one hold the slide over the flame for long duration, one would incinerate the cells.

Thus, heat fixing is important for the proper attachment of the bacterial cells with the slide and for proper visibility of the smear. One should also be careful about the factors, which may lead to  poor results.

Find out more information about heat fixation here:

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