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The usual purpose is to reveal cytological details that might otherwise not be apparent; however, staining can also reveal where certain chemicals or specific chemical reactions are taking place within cells or tissues. In vitro staining involves colouring cells or structures that have been removed from their biological context. Certain stains ...
The chemical composition and pH value of the buffer solution also contribute to the effectiveness of heat-induced antigen retrieval. [1] Thus, the AR-immunohistochemistry protocol must be optimized for each tissue type, fixation method, and antigen using a "test battery" to maximize antigen recovery in formalin fixed paraffin embedded sections. [1]
Heat fixation is used for the fixation of single cell organisms, most commonly bacteria and archaea. The organisms are typically mixed with water or physiological saline which helps to evenly spread out the sample. Once diluted, the sample is spread onto a microscope slide. This diluted bacteria sample is commonly referred to as a smear after ...
Immunohistochemistry can be performed on tissue that has been fixed and embedded in paraffin, but also cryopreservated (frozen) tissue.Based on the way the tissue is preserved, there are different steps to prepare the tissue for immunohistochemistry, but the general method includes proper fixation, antigen retrieval incubation with primary antibody, then incubation with secondary antibody.
One of the main difficulties with IHC staining is overcoming specific or non-specific background. Optimisation of fixation methods and times, pre-treatment with blocking agents, incubating antibodies with high salt, and optimising post-antibody wash buffers and wash times are all important for obtaining high quality immunostaining.
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