Immunohistochemical Techniques

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Immunohistochemistry is a technique that involves the use of antibody-antigen interactions in order to identify cellular and tissue constituents. One way this can be done by labelling known antibodies with enzymes which produce a coloured product after reacting and then monitoring the sample to see whether a reaction has taken place. In immunohistochemistry, the preservation of the antigenic determinants, also known as epitopes, and binding sites is vital in order to ensure that an accurate result is obtained. However, tissue samples need to go through various processes in a pathology laboratory before they are ready for testing and some of these processes can alter the structure of the epitopes so it’s important to consider this when choosing reagents in order to reduce the risk of this happening.
Tissue samples need to go through fixation before any tests can be carried out on them. This is to ensure that the tissue samples are well preserved; preventing the loss of molecular structure and cellular morphology, to stop autolysis and ensuring any pathogens residing in the tissue are destroyed. The major objective to remember when choosing a fixative is that it must maintain clear and consistent morphological features within the tissues for viewing with a microscope and because of this the most widely used method of fixation is to use 10% neutral buffered formalin as it provides a good view of the morphology within the tissues. Formalin is an aqueous solution that contains approximately 4% formaldehyde and the formaldehyde reacts with itself to produce polymers and methylene bridges which fix the tissues by creating cross links between the proteins in the tissues, and other molecules, such as carbohydrates and fats, are trapped bet...

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