What stains are used in immunohistochemistry?
Common counterstains include hematoxylin, eosin, nuclear fast red, methyl green, DAPI, and Hoechst fluorescent stain. The following representative example, Hoechst fluorescent dye was used as a counterstain for IHC detection of the protein, vimentin.
How does immunohistochemistry staining work?
Immunohistochemical staining is accomplished with antibodies that recognize the target antigen. Since antibodies are highly specific, the antibody will bind only to the antigen of interest in the tissue section. The antibody-antigen interaction is then visualized using different detection systems.
What are the different detection methods for IHC?
Various Detection Methods for IHC. Choosing the Right Reporter Enzyme and Chromogen. Multiplexing Chromogenic IHC. Common Counterstains for IHC.
Which method of immunohistochemistry is more sensitive?
Indirect Method This method is more sensitive due to signal amplification through several secondary antibody reactions with different antigenic sites on the primary antibody.
What is background staining in immunohistochemistry?
Background staining is thought to occur as a result of either non-specific antibody (Ab) binding to endogenous Fc receptors (FcRs) or a combination of ionic and hydrophobic interactions.
What is the principle of immunochemistry?
Immunohistochemistry (IHC) is a method for detecting antigens or haptens in cells of a tissue section by exploiting the principle of antibodies binding specifically to antigens in biological tissues. The antibody-antigen binding can be visualized in different manners.
What is HRP polymer?
VisUCyte™ HRP Polymer is a Biotin-free detection reagent that overcomes problems associated with Avidin-Biotin detection chemistry, such as the extra quenching steps required to inhibit endogenous Biotin and Avidin staining.
What is the amplification in immunohistochemical detection?
Signal amplification techniques greatly enhance the sensitivity of immunohistochemical (IHC) and in situ hybridization (ISH) methods. In particular, catalyzed signal amplification (CSA) using labeled tyramide or Nanogold-silver staining is an important signal amplification tool.
What does H and E stand for?
The hematoxylin and eosin stain (H&E) is the most widely used stain in histology and histopathology laboratories.