According to the biotechnology experiment summary, we know that immunohistochemistry is the use of labeled specific antibodies for tissue and cell in situ qualitative, localized or quantitative studies on the distribution and content of certain chemical components in tissue sections or cell specimens. It is an immunocytochemical technique. The following is a brief introduction to several commonly used immunohistochemical techniques, and hopes to deepen our understanding of immunohistochemistry.
Several commonly used immunohistochemical techniques are available in the following methods: :
(1) Immunofluorescence method: It is the earliest established immunohistochemical technique. It uses the principle of antigen-antibody specific binding, first labeled known antibodies with fluorescein, as a probe to examine the corresponding antigen in cells or tissues, and observed under a fluorescence microscope. When the fluorescein in the antigen-antibody complex is irradiated with the excitation light, a certain wavelength of fluorescence is emitted, thereby determining the localization of an antigen in the tissue, and further quantitative analysis. Because of its strong specificity, high sensitivity, fast and simple, immunofluorescence technology is widely used in clinical pathology diagnosis and testing.
(2) Immunoenzyme labeling method: The immunoenzymatic labeling method is a technique developed in the 1960s following immunofluorescence. The basic principle is that the enzyme-labeled antibody acts on the tissue or cells, and then the substrate of the enzyme is added to form a colored insoluble product or a particle having a certain electron density, and the surface of the cell and various cells are irradiated by light microscopy or electron microscopy. The antigen component was subjected to localization studies. Immunolabeling technology is currently the most commonly used technology. The main advantages of this method compared with immunofluorescence technology are: accurate positioning, good contrast, long-term preservation of stained specimens, suitable for light and electron microscope research. The development of immunolabeling methods has been very rapid, and a variety of labeling methods have been derived. With the continuous improvement and innovation of the methods, the specificity and sensitivity are continuously improved, and the use is more and more convenient. At present, ABC method, SP three-step method, and ready-to-use two-step detection system are widely used in pathological diagnosis.
(3) Immunocolloidal gold technology: The immunocolloidal gold technique uses a special metal particle such as colloidal gold as a marker. Colloidal gold refers to a gold hydrosol that adsorbs proteins quickly and steadily without a significant effect on the biological activity of the protein. Therefore, using a colloidal gold-labeled primary antibody, secondary antibody, or other molecule that specifically binds to an immunoglobulin (such as Staphylococcal Protein A) can be used as a probe to characterize, localize, or even quantify antigens in tissues or cells. the study. Because colloidal gold has particles of different sizes and the electron density of colloidal gold is high, the immunocolloidal gold technique is particularly suitable for single-label or multi-label localization studies of immunoelectron microscopy. Since colloidal gold itself is light to deep red, it is also suitable for light microscopy. For example, the silver-enhanced immunogold and silver rule is more convenient for light microscopy.
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