When Tissue Antigens and Antibodies Get Along

Abstract

Once focused mainly on the characterization of neoplasms, immunohistochemistry (IHC) today is used in the investigation of a broad range of disease processes with applications in diagnosis, prognostication, therapeutic decisions to tailor treatment to an individual patient, and investigations into the pathogenesis of disease. This review addresses the technical aspects of immunohistochemistry (and, to a lesser extent, immunocytochemistry) with attention to the antigen-antibody reaction, optimal fixation techniques, tissue processing considerations, antigen retrieval methods, detection systems, selection and use of an autostainer, standardization and validation of IHC tests, preparation of proper tissue and reagent controls, tissue microarrays and other high-throughput systems, quality assurance/quality control measures, interpretation of the IHC reaction, and reporting of results. It is now more important than ever, with these sophisticated applications, to standardize the entire IHC process from tissue collection through interpretation and reporting to minimize variability among laboratories and to facilitate quantification and interlaboratory comparison of IHC results.

Predictive value of BRCA1 expression on the efficacy of chemotherapy based on anti-microtubule agents: a pooled analysis across different malignancies and agents

“…Since significant heterogeneity was observed in the overall analyses, we carried out additional subgroup analyses. Interestingly, our results show that using non-immunohistochemical (PCR and Relative cDNA quantification) detection methods offer the notable result that high BRCA1 expression was associated with higher sensitivity of anti-microtubule agents whereas the difference in the subgroup using IHC was not significant (for non-immunohistochemical study, high vs. IHCis a process that exploits the principle of antibodies binding specifically to antigens in biological tissues to detect antigens (e.g., proteins) (28). The detection target of IHC is the proteins which are at the last destination to take a leading role in biological effects, so it has clinical significance but it also carries obvious limitations.…”

SlideToolkit: An Assistive Toolset for the Histological Quantification of Whole Slide Images

“…For immunohistochemical staining, antibodies are visualized by a chromogenic substrate, such as DAB (brown), 3-AEC (red) or a fluorescent dye. [2] To visualize the remaining tissue a hematoxylin (blue) counter stain is often applied. Until recently, we applied manual semi-quantitative scoring methods or case-by-case quantitative scoring of immunohistochemically stained cross-sections.…”

Genetic mechanisms regulating proliferation and cell specification in the Drosophila embryonic CNS

“…This technique is widely used since 1941 to identify e.g., the localization and distribution of biological markers, and the variation of their expression in different cell types [207]. Visualization of the antibody is commonly done by enzymatic reaction or different fluorophore tags i.e., immunofluorescence [208]. IHC approach is used in all of the papers of this thesis.…”

“…The primary antibodies, which are tested for validity in mutant backgrounds, are added to the tissue, and subsequently visualized by fluorophore-tagged secondary antibodies (as mentioned in details within each paper). [208].…”