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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.
Immunohistochemistry or IHC staining of tissue sections (or immunocytochemistry, which is the staining of cells), is perhaps the most commonly applied immunostaining technique. [2] While the first cases of IHC staining used fluorescent dyes (see immunofluorescence ), other non-fluorescent methods using enzymes such as peroxidase (see ...
In contrast, immunohistochemical samples are sections of biological tissue, where each cell is surrounded by tissue architecture and other cells normally found in the intact tissue. Immunocytochemistry is a technique used to assess the presence of a specific protein or antigen in cells (cultured cells, cell suspensions) by use of a specific ...
After the development of AR, enzyme digestion was rarely used in immunohistochemistry for formalin fixed paraffin embedded tissue sections. Although enzyme digestion and antigen retrieval target the same problem in immunohistochemistry, these two techniques differs in both the mode of action and effectiveness, warranting distinct nomenclature. [8]
Immunohistochemistry and immunocytochemistry are methods used to determine in which cells or parts of cells, a particular protein or other macromolecule are located. These stains use antibodies to bind to specific antigens, usually of protein or glycoprotein origin. Since antibodies are normally invisible, special strategies must be employed to ...
Each microarray block can be cut into 100 – 500 sections, which can be subjected to independent tests. Tests commonly employed in tissue microarray include immunohistochemistry, and fluorescent in situ hybridization. Tissue microarrays are particularly useful in analysis of cancer samples. One variation is a Frozen tissue array.
Therefore, the choice of fixative and fixation protocol may depend on the additional processing steps and final analyses that are planned. For example, immunohistochemistry uses antibodies that bind to a specific protein target. Prolonged fixation can chemically mask these targets and prevent antibody binding.
Use of alcian blue has historically been a popular staining method in histology especially for light microscopy in paraffin embedded sections and in semithin resin sections. The tissue parts that specifically stain by this dye become blue to bluish-green after staining and are called "Alcianophilic" (comparable to "eosinophilic" or "sudanophilic").