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Immunocytochemistry and Immunohistochemistry

Immunocytochemistry and Immunohistochemistry

Immunocytochemistry (ICC) and immunohistochemistry (IHC) are techniques that use antibodies to check for specific proteins or antigens in a sample. The technique was first published by Albert Coons in 1941 to detect the presence of pneumococcal antigen in tissue sections from mice infected with Pneumococcus. Immunocytochemistry helps localization of proteins or antigens in individual cells like blood cells, stem cells, etc., while immunohistochemistry does the same for tissue samples.
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Updated: May 5, 2026

Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging

Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging

Published on: April 8, 2016

Practical quantification of immunohistochemistry antigen concentrations and reaction-diffusion parameters.

Woody Perng¹, Berenice Mbiribindi², Benjamin Thomas Andrews³

¹Research Pathology, Genentech, Inc., South San Francisco, CA 94080, USA.

Biorxiv : the Preprint Server for Biology

|May 4, 2026

Summary

This study introduces a new method to quantify immunohistochemistry (IHC) by modeling reaction-diffusion kinetics. This allows for more accurate measurement of antigen concentration in FFPE samples, improving diagnostic accuracy.

Keywords:

Immunohistochemistry effective diffusion coefficient reaction-diffusion

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Last Updated: May 5, 2026

Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging

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Published on: April 8, 2016

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Published on: January 7, 2019

Area of Science:

Biomedical research
Diagnostic medicine
Biophysics

Background:

Immunohistochemistry (IHC) is widely used but lacks theoretical tools for interpretation.
Current IHC protocols are often empirically optimized, limiting quantitative accuracy.
Stain intensity is not directly interpretable as chemical antigen concentration.

Purpose of the Study:

To develop practical theoretical and experimental tools for quantitative IHC analysis.
To enable measurement of both antigen concentration and reaction-diffusion parameters in FFPE samples.
To improve the objectivity and reliability of IHC interpretation.

Main Methods:

Developed a fast interpolation method to model IHC reaction-diffusion behavior.
Established experimental methods to characterize IHC kinetic parameters in FFPE samples.
Integrated modeling and experimental methods for quantitative analysis.

Main Results:

Direct immunofluorescent detection shows nanomolar sensitivity and a >1000-fold dynamic range.
Antibody diffusion in FFPE samples can be >1000-fold slower than in aqueous solutions.
Diffusion-limited conditions can affect IHC reaction time courses and antigen concentration interpretation.

Conclusions:

The developed framework advances IHC from qualitative to quantitative analysis.
Enables objective interpretation of IHC results across different conditions and labs.
Allows IHC staining to be interpreted as molar concentrations, improving biological correlation analysis.