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Updated: Jul 20, 2026
Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging
Published on: April 8, 2016
Quantification of immunohistochemistry--issues concerning methods, utility and semiquantitative assessment II
1Department of Pathology, Keck School of Medicine of the University of Southern California, Los Angeles, CA 90033, USA. taylor@pathfinder.usc.edu
Insights
Immunohistochemistry (IHC) has evolved for tumor diagnosis and prognosis. This review examines IHC
Area of Science:
- Pathology
- Biotechnology
- Cancer Research
Background:
- Immunohistochemistry (IHC) has been a cornerstone of diagnostic surgical pathology for decades.
- Initially used as qualitative 'special stains' for tumor classification, IHC has evolved significantly.
- The rise of molecular biology has shifted focus to prognostic markers, demanding quantitative analysis.
Purpose of the Study:
- To review the evolution of immunohistochemistry (IHC) in diagnostic surgical pathology.
- To explore the limitations of semi-quantitative IHC methods.
- To discuss the potential of IHC as a true quantitative immunoassay in tissue sections.
Main Methods:
- Review of historical and current applications of immunohistochemistry (IHC).
- Analysis of the transition from qualitative to quantitative IHC.
- Exploration of the challenges and possibilities in achieving quantitative IHC.
Main Results:
- Immunohistochemistry (IHC) has transitioned from qualitative assessment to a need for quantitative analysis.
- Current semi-quantitative IHC methods have inherent limitations.
- There is a growing demand for precise quantification of markers in tumor cells.
Conclusions:
- Immunohistochemistry (IHC) is a critical tool in pathology, but its full potential requires quantitative accuracy.
- Overcoming the limitations of semi-quantitative methods is essential for advancing diagnostic and prognostic capabilities.
- Developing true quantitative IHC assays for tissue sections is a key future direction.
Abstract:
Immunohistochemistry is entering its fourth decade of use on formalin-fixed paraffin-embedded tissues. Over this period the method has evolved to become a major part of the practice of diagnostic surgical pathology worldwide. From the beginning immunohistochemistry has been adapted to provide a range of markers of cell lineage and tissue type, with particular application to the diagnosis and classification of tumours. In this modality immunohistochemical methods were employed simply as 'special stains', the results of which were evaluated qualitatively by the pathologist, as for any other stain. More recently, attention has shifted to the demonstration of prognostic markers in tumour cells, driven by the advent of molecular biology and the discovery of numerous regulatory molecules, coupled with manufacture of the corresponding specific antibodies. Immunohistochemistry has rapidly adapted to this new use, but in so doing the demand for quantification has become paramount; it is no longer enough that the 'stain' is there; rather it is a question of 'How much is there?'. This review explores the limitations of immunohistochemistry when employed in a semiquantitative mode, and explores the possibility of fulfilling the full potential of immunohistochemistry, as a true quantitative immunoassay applied in a tissue section environment.
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