{"id":830,"date":"2018-06-10T20:51:08","date_gmt":"2018-06-11T01:51:08","guid":{"rendered":"http:\/\/www.bocsci.com\/blog\/?p=830"},"modified":"2018-06-10T20:51:08","modified_gmt":"2018-06-11T01:51:08","slug":"what-is-immunocytochemistry","status":"publish","type":"post","link":"https:\/\/www.bocsci.com\/blog\/what-is-immunocytochemistry\/","title":{"rendered":"What Is Immunocytochemistry?"},"content":{"rendered":"

Immunocytochemistry<\/strong><\/p>\n

Immunocytochemistry is the use of antibodies for identifying proteins and\u00a0molecules in cells and\u00a0tissues viewed under a microscope. Immunocytochemistry\u00a0harnesses the power of antibodies to\u00a0give highly specific binding to unique\u00a0sequences of amino acids in proteins. Perhaps the most\u00a0exciting part of using\u00a0antibodies is that new antibodies can be generated on an as-needed basis,\u00a0thus\u00a0providing a constant source of new reagents. Scientists are constantly generating\u00a0new\u00a0antibodies to specific parts of molecules thus driving continual evolution of\u00a0immunocytochemistry. Identifying the location of antibodies in cells is based on\u00a0availability of\u00a0labels that is, itself, rapidly advancing. As time passes, immunocytochemistry continues to\u00a0respond to new development of labels and advanced methods\u00a0of labeling molecules.<\/p>\n

If the terms immunocytochemistry and immunohistochemistry seem similar\u00a0then here is why.\u00a0Many years ago, immunocytochemistry was defined as the\u00a0use of antibodies to study cells in the form of cultures or smears from animals.\u00a0Immunohistochemistry, on the other hand, was defined\u00a0as the use of antibodies to\u00a0study paraffin sections from human tissue. Today, immunohistochemistry is still the use of antibodies in paraffin sections in human pathology, but\u00a0the definition of\u00a0immunocytochemistry has changed. Immunocytochemistry is the use of<\/i><\/em>\u00a0<\/i><\/em>antibodies<\/i><\/em>\u00a0<\/i><\/em>in animal research with cells and tissues fixed in paraformaldehyde<\/i><\/em>.<\/p>\n

This new definition of immunocytochemistry derives from advances in antibodylabeling methods\u00a0in recent years. These advances resulted from specific needs in\u00a0animal research. Initially, formalin-fixed paraffin sections were used for immunohistochemistry; however, results were\u00a0inconsistent. In most cases, the antibody\u00a0did not label anything or it labeled too many cells and was dubbed \u201cover fixed.\u201d\u00a0This problem led to the development of the epitope retrieval or antigen retrieval\u00a0methods, where sections of tissue are treated with heat in buffers before antibody\u00a0incubations. Unfortunately, epitope retrieval methods can be unique from antibody\u00a0to antibody and also, for the same antibody, from tissue to tissue. Epitope retrieval\u00a0is complicated and best avoided. For animal research, a simple method was then\u00a0developed where tissue was fixed in paraformaldehyde and not formalin or alcohol\u00a0and subsequently frozen sections were cut on a\u00a0cryostat. This eliminated the steps\u00a0of dehydration, embedding in paraffin, rehydration after\u00a0sectioning, and epitope\u00a0retrieval before antibody incubation. This was a major breakthrough.<\/p>\n

What Can Immunocytochemistry Tell Us?<\/p>\n

Immunocytochemistry<\/i><\/em>\u00a0harnesses antibodies that are specific reagents and which\u00a0allow unique\u00a0detection of proteins and molecules. Using antibodies requires specific methods, labels, and\u00a0controls. Performing immunocytochemistry experiments\u00a0requires some basic knowledge of biology.<\/p>\n

Much of the data collected in biomedical research today results from biochemical and molecular\u00a0methods, where many cells are pooled for analysis. For example,\u00a0enzyme assay of the liver will give values that, when repeated, should be statistical similar and should provide reliable average\u00a0values with standard errors. When\u00a0this and similar methods pool many cells for analysis, they are broadly defined as\u00a0\u201cpopulation studies\u201d. However, problems result, because not all liver\u00a0cells might have the specific enzyme of interest. So changes found with the enzyme\u00a0assay might be\u00a0due to enzyme activity in all of the liver cells or might be due to\u00a0enzyme activity in only some of\u00a0the cells. Rather than assuming all of the cells in\u00a0the liver have the enzyme, the complementary\u00a0approach is to look at the cells with\u00a0morphological methods.<\/p>\n

Morphological approaches in biomedical research can include a wide range\u00a0of microscopes, but today typically employ immunocytochemistry that can give\u00a0us information about individual liver\u00a0cells containing the specific enzyme.\u00a0Immunocytochemistry uses antibodies to bind proteins and labels to show protein\u2019s\u00a0location. If, for example, the enzyme is a marker for inflammation, then\u00a0the location\u00a0of cells with this enzyme tells us which cell types have the inflammatory response.\u00a0Thus, immunocytochemistry methods are broadly defined as \u201cindividual studies\u201d\u00a0of\u00a0single cells or cell groups. The resulting data tell us about location of the enzyme.<\/p>\n

Reference:<\/p>\n

Burry, R. W. Immunocytochemistry. A Practical Guide for Biomedical Research. 2010.<\/p>\n","protected":false},"excerpt":{"rendered":"

Immunocytochemistry Immunocytochemistry is the use of antibodies for identifying proteins and\u00a0molecules in cells and\u00a0tissues viewed under a microscope. Immunocytochemistry\u00a0harnesses the power of antibodies to\u00a0give highly specific binding to unique\u00a0sequences of […]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[147],"tags":[],"_links":{"self":[{"href":"https:\/\/www.bocsci.com\/blog\/wp-json\/wp\/v2\/posts\/830"}],"collection":[{"href":"https:\/\/www.bocsci.com\/blog\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.bocsci.com\/blog\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.bocsci.com\/blog\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.bocsci.com\/blog\/wp-json\/wp\/v2\/comments?post=830"}],"version-history":[{"count":1,"href":"https:\/\/www.bocsci.com\/blog\/wp-json\/wp\/v2\/posts\/830\/revisions"}],"predecessor-version":[{"id":831,"href":"https:\/\/www.bocsci.com\/blog\/wp-json\/wp\/v2\/posts\/830\/revisions\/831"}],"wp:attachment":[{"href":"https:\/\/www.bocsci.com\/blog\/wp-json\/wp\/v2\/media?parent=830"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.bocsci.com\/blog\/wp-json\/wp\/v2\/categories?post=830"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.bocsci.com\/blog\/wp-json\/wp\/v2\/tags?post=830"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}