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Immunohistochemistry for Pathologists: Protocols, Pitfalls, and Tips
So-Woon Kim1, Jin Roh1, Chan-Sik Park1
1Department of Pathology, Asan Medical Center, University of Ulsan College of Medicine, 88 Olympic-ro, 43-gil, Songpa-gu, Seoul 05505, Korea.
Insights
This review offers practical tips and troubleshooting advice for immunohistochemistry (IHC), a key diagnostic tool for pathologists. It covers IHC procedures, interpretation, and analysis to improve accuracy in research and clinical practice.
Area of Science:
- Pathology
- Biomedical Research
- Molecular Diagnostics
Background:
- Immunohistochemistry (IHC) is crucial for pathologists in diagnostics and research, enabling confirmation of target molecule expression within the microenvironment.
- Despite advancements in automation and standardization, optimizing IHC protocols and ensuring accurate interpretation remain critical challenges.
- This review addresses common pitfalls and provides practical guidance for IHC application.
Approach:
- Summarizes the general IHC procedure, detailing potential pitfalls and offering solutions for each step.
- Includes a comprehensive troubleshooting guide for common IHC issues.
- Discusses accurate interpretation methods, including quantification and analysis techniques for IHC results.
Key Points:
- Highlights critical considerations for IHC optimization and interpretation.
- Provides actionable tips for practicing pathologists and residents.
- Focuses on enhancing the reliability and reproducibility of IHC staining.
Conclusions:
- Aims to serve as a foundational reference for improving IHC practice and publication quality.
- Emphasizes the importance of meticulous protocol execution and interpretation for diagnostic and research utility.
- Offers strategies to overcome common challenges in immunohistochemistry workflows.
Abstract:
Immunohistochemistry (IHC) is an important auxiliary method for pathologists in routine diagnostic work as well as in basic and clinical research including exploration of biomarkers, as IHC allows confirmation of target molecule expressions in the context of microenvironment. Although there has been a considerable progress in automation and standardization of IHC, there are still many things to be considered in proper optimization and appropriate interpretation. In this review, we aim to provide possible pitfalls and useful tips for practicing pathologists and residents in pathology training. First, general procedure of IHC is summarized, followed by pitfalls and tips in each step and a summary of troubleshooting. Second, ways to an accurate interpretation of IHC are discussed, with introduction to general quantification and analysis methods. This review is not intended to provide complete information on IHC, but to be used as a basic reference for practice and publication.
