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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.
Heat-induced antigen retrieval is the most widely used pretreatment in immunohistochemistry for formalin fixed paraffin embedded tissue sections. It requires boiling deparaffinized formalin fixed paraffin embedded tissue sections in either water or various buffer solutions.
Tissue preparation or fixation is essential for the preservation of cell morphology and tissue architecture. Inappropriate or prolonged fixation may significantly diminish the antibody binding capability. Many antigens can be successfully demonstrated in formalin-fixed paraffin-embedded tissue sections. However, some antigens will not survive ...
In the tissue microarray technique, a hollow needle is used to remove tissue cores as small as 0.6 mm in diameter from regions of interest in paraffin-embedded tissues such as clinical biopsies or tumor samples. These tissue cores are then inserted in a recipient paraffin block in a precisely spaced, array pattern.
Tissue embedded within optimal cutting temperature compound (OCT), mounted on a chuck in a cryostat, and ready for section production. The frozen section procedure is a pathological laboratory procedure to perform rapid microscopic analysis of a specimen. It is used most often in oncological surgery. [1] The technical name for this procedure is ...
The substance used to embed tissue is embedding media, which is chosen depends on the category of the microscope, category of the micro tome, and category of tissue. [23] Paraffin wax, whose melting point is from 56 to 62°C, is commonly used for embedding. [22] Tissue processing - Tissue sections on slides are stained on an automated stainer
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").
Paraffin wax may also be too soft in relation to the tissue, the heat of the melted wax may alter the tissue in undesirable ways, or the dehydrating or clearing chemicals may harm the tissue. [12] Alternatives to paraffin wax include, epoxy , acrylic , agar , gelatin , celloidin , and other types of waxes.