Technical Aspects of Immunohistochemistry

Abstract

Abstract.Immunohistochemistry is an integral technique in many veterinary laboratories for diagnostic and research purposes. In the last decade, the ability to detect antigens (Ags) in tissue sections has improved dramatically, mainly by countering the deleterious effects of formaldehyde with antigen retrieval (AR) and increasing sensitivity of the detection systems. In this review, I address these topics and provide an overview of technical aspects of immunohistochemistry, including those related to antibodies (Abs) and Ags, fixation, AR, detection methods, background, and troubleshooting. Microarray technology and the use of rabbit monoclonal Abs in immunohistochemistry are also discussed.

The free fatty acid metabolome in cerebral ischemia following human mesenchymal stem cell transplantation in rats

“…Immunohistochemistry is the localization of antigens or proteins in cells of a tissue section exploiting the principle of antigen-antibody interactions that are visualized by a marker in biological tissues [30][31][32]. Animals were sacrificed 5 days post MCAo.…”

Morphological Distinction of Histiocytic Sarcoma from Other Tumor Types in Bernese Mountain Dogs and Flatcoated Retrievers

“…Absence, weak or variable IHC staining intensity can be caused by the formaldehyde of the FFPE procedure. Formaldehyde makes recognition of antigens difficult, and overfixation with this preserving agent can result in detection failure (or only weak reaction) (23). Differences in the degree of differentiation of the histiocytic tumor cells can result in different intensity of staining within and between cases (17).…”

A comparison of immunohistochemistry and in situ hybridization for the detection of porcine circovirus type 2 in pigs

“…The correct interpretation of IHC results may be hampered by the presence of pigments, such as ferrous pigment, for example hemosiderin, which can reduce the signal-to noise ratio (Ramos-Vara 2005). To avoid confusion in the interpretation of such findings the recommended procedure is additional treatment of tissue with a 1% solution of dithionite in pH 5.0 acetate buffer or using a detection system producing a different colored precipitate (Ramos-Vara 2005).…”

“…To avoid confusion in the interpretation of such findings the recommended procedure is additional treatment of tissue with a 1% solution of dithionite in pH 5.0 acetate buffer or using a detection system producing a different colored precipitate (Ramos-Vara 2005). In IHC protocols using diaminobenzidine (DAB) as a chromogen, giving brown staining, the presence of iron deposits may be mistaken for positive staining.…”