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Updated: Apr 5, 2026
Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry
Published on: July 26, 2019
A Dermatopathologist's Guide to Troubleshooting Immunohistochemistry--Part 2: Troubleshooting Immunohistochemical
Amanda F Marsch1, Jonathan N Truong, Melissa M McPherson
1*Dermatology Resident, Department of Dermatology, University of Illinois at Chicago, Chicago, IL; and †Laboratory Manager (J.N.T.), IHC HistoTechnician (M.M.M.), Dermatopathologist (J.M.J.-H.), Director (D.M.E.), Ackerman Academy of Dermatopathology, University of Illinois, Chicago, IL.
Insights
Immunohistochemistry (IHC) staining requires careful attention to detail to avoid errors. Understanding potential pitfalls in IHC methodology is crucial for accurate tumor diagnosis and therapeutic target identification.
Area of Science:
- Pathology
- Laboratory Medicine
- Oncology
Background:
- Immunohistochemistry (IHC) is a vital diagnostic tool in pathology.
- Accurate IHC staining is essential for differentiating benign from malignant tumors and identifying therapeutic targets.
- Previous installments in this series have addressed some IHC procedures.
Purpose of the Study:
- To highlight potential sources of error in immunohistochemistry (IHC) staining.
- To emphasize the importance of understanding IHC methodology pitfalls for accurate interpretation.
- To guide pathologists in troubleshooting and avoiding false results in IHC.
Main Methods:
- Discussion of common issues encountered during IHC procedures.
- Review of critical steps in IHC staining relevant to potential errors.
- Case examples or theoretical scenarios illustrating staining pattern variations.
Main Results:
- Unexpected staining patterns in IHC can result from errors at various procedural stages.
- Familiarity with IHC methodology is key to identifying and rectifying staining anomalies.
- Potential for false-positive or false-negative results necessitates careful interpretation.
Conclusions:
- Pathologists must possess a deep understanding of IHC pitfalls for reliable laboratory troubleshooting.
- Accurate interpretation of IHC staining is paramount to avoid diagnostic errors.
- Mastery of IHC techniques ensures correct patient diagnosis and treatment planning.
Abstract:
Unexpected staining patterns can arise from problems occurring in any of the steps required for IHC, some of which are discussed in part I of this CME series. Whether used to differentiate benign from malignant tumors, identify tumor subtypes, subtypes of hematopoietic malignancies, or identifying targets for therapy, the pathologist must be intimately familiar with the potential pitfalls that are inherent in the IHC methodology to troubleshoot problems in the laboratory, and more importantly, when interpreting immunohistochemical staining, to avoid pitfalls of false-positive or false-negative stains.
