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Published on: November 14, 2016
Fixation and epitope retrieval in diagnostic immunohistochemistry: a concise review with practical considerations
R T Miller1, P E Swanson, M R Wick
1Immunohistochemistry Division, ProPath Laboratory, Inc, Dallas, Texas 75247, USA. rmiller@propathlab.com
Insights
Diagnostic immunohistochemistry (DIHC) significantly impacts surgical pathology. Optimizing laboratory variables, particularly tissue processing for paraffin sections, is crucial for quality assurance and reliable results in DIHC.
Area of Science:
- Pathology
- Laboratory Medicine
- Histotechnology
Background:
- Diagnostic immunohistochemistry (DIHC) has transformed surgical pathology practice over 25 years.
- Methodological variations in DIHC significantly influence diagnostic outcomes.
- Paraffin section-based immunohistology is the most prevalent adjunctive morphologic procedure (AMP) in hospital pathology.
Purpose of the Study:
- To review key variables affecting DIHC results.
- To provide guidance on optimizing these variables for quality assurance in DIHC laboratories.
- To highlight the importance of fixation and processing for optimal AMP performance.
Main Methods:
- Review of methodological variables in diagnostic immunohistochemistry.
- Focus on processing factors impacting paraffin section-based immunohistology.
- Inclusion of considerations for other tissue analyses.
Main Results:
- Identified key processing variables that influence DIHC outcomes.
- Demonstrated how optimizing these variables enhances quality assurance.
- Provided insights into best practices for fixation and tissue processing.
Conclusions:
- Effective quality assurance in DIHC relies on meticulous control of methodological details.
- Optimized tissue processing is essential for accurate and reproducible DIHC results.
- The principles discussed apply to paraffin-section immunohistology and other tissue analyses.
Abstract:
Diagnostic immunohistochemistry (DIHC) is an extremely valuable technological development that has revolutionized the practice of surgical pathology over the past 25 years. Like any laboratory procedure, however, the precise concatenation of methodological details that are used by any given investigator has a major effect on final results. In this overview, the authors consider several of these variables and illustrate how they may be optimized in an effective and efficient approach to quality assurance in the DIHC laboratory. The great majority of this discussion concerns processing factors as they affect paraffin section-based immunohistology because that is still, by far, the most common adjunctive morphologic procedure (AMP) used in hospital pathology. However, information is also presented on elements of tissue processing that affect other analyses as well. The material presented here is a direct reflection of the conjoint experiences of the authors. Although we discuss our preferences regarding many technical aspects of this discipline, we certainly do not mean to imply that ours are the only methods that should be utilized. Nevertheless, it is hoped that they may serve to crystallize basic concepts surrounding fixation, processing, and optimal performance of AMPs.
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