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Pitfalls Caveats in Chromogenic Immunostaining
In daily diagnostic pathology services, chromogenic immunostaining using formalin-fixed, paraffin-embedded (FFPE) sections is indispensable to help in the histopathological diagnosis fundamentally based on hematoxylin and eosin (H&E) staining. Differential diagnoses are determined first, and appropriate immunohistochemical markers for the diagnoses are then selected. The expression of lymphocyte surface markers, hormones and tumor markers contributes to the functional classification of tumors. The degree of malignancy can be analyzed by immunostaining for p53 and Ki-67 (MIB-1), a cell proliferation marker, in a variety of malignancies including carcinomas of the oral cavity, colon, and breast.
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.
2. Techniques of Immunohistochemistry
Techniques of immunohistochemistry (IHC) or immunostaining have already been established . Particularly 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 herein were obtained by the author and/or the author’s colleagues.
3. The Need for Chromogenic Immunostaining in Diagnostic Pathology
4. Selection of Methodology for Chromogenic Immunostaining
The entry is from 10.3390/cells10061501
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