Abstract
Chromogenic immunohistochemistry (immunostaining using an enzyme-labeled probe) is an essential histochemical technique for analyzing pathogenesis and making a histopathological diagnosis in routine pathology services. In neoplastic lesions, immunohistochemistry allows the study of specific clinical and biological features such as histogenesis, behavioral characteristics, therapeutic targets, and prognostic biomarkers. The needs for appropriate and reproducible methods of immunostaining are prompted by technical development and refinement, commercial availability of a variety of antibodies, advanced applicability of immunohistochemical markers, accelerated analysis of clinicopathological correlations, progress in molecular targeted therapy, and the expectation of advanced histopathological diagnosis. However, immunostaining does have various pitfalls and caveats. Pathologists should learn from previous mistakes and failures and from results indicating false positivity and false negativity. The present review article describes various devices, technical hints, and trouble-shooting guides to keep in mind when performing immunostaining.
Highlights
Techniques of immunohistochemistry (IHC) or immunostaining have already been established [1,2,3,4,5,6,7,8]
We evaluated the localization of CD4, interleukin-6 (IL-6), and interferon-gamma (IFN-γ) in heat-retrieved FFPE sections of pharyngeal tonsil with the catalyzed signal amplification (CSA)-II system
When Unexpected Results Are Obtained Clinicopathological significance should be recognized when immunohistochemical results are different from hematoxylin and eosin (H&E)-based expectations
Summary
Techniques of immunohistochemistry (IHC) or immunostaining have already been established [1,2,3,4,5,6,7,8]. For diagnostic IHC, chromogenic IHC (immunostaining using enzyme-labeled probes) is commonly utilized. Measures and tools to prevent technical artifacts and appropriate trouble-shooting tips are needed. The present review article provides an overview of technical aspects, knowhow, pitfalls, and trouble-shooting guides for routinely performing chromogenic immunostaining, especially for experienced personnel with immunohistochemical expertise. It should be emphasized that all data presented were obtained by the author and/or the author’s colleagues. 2. The Need for Chromogenic Immunostaining in Diagnostic Pathology IHC is critical both to informing the biology of lesions and to formulating the histopathological diagnosis. The patient’s prognosis and response to therapeutic approaches may be predicted
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