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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.
These...

Immunogold Electron Microscopy

Immunogold Electron Microscopy

Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.

Immunofluorescence Microscopy

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.

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Related Experiment Video

Updated: Oct 13, 2025

Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections

Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections

Published on: November 14, 2016

Immunochemistry Analysis Using Chromogenic Substrates on Tissue Sections.

Preethi Vijayaraj¹

¹Immunology and Respiratory Research, Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, CT, USA. preethi.vijayaraj@boehringer-ingelheim.com.

Methods in Molecular Biology (Clifton, N.J.)

|November 12, 2021

Summary

Immunohistochemistry (IHC) visualizes protein abundance and distribution using antigen-antibody reactions. This guide details standard IHC protocols with chromogenic substrates, including tissue pre-treatment and troubleshooting.

Keywords:

Antigen retrieval Blocking Chromogenic substrate Detection Fixation Immunohistochemistry Permeabilization Troubleshooting

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Histological-Based Stainings Using Free-Floating Tissue Sections

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Last Updated: Oct 13, 2025

Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections

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Histological-Based Stainings Using Free-Floating Tissue Sections

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Published on: August 25, 2020

Area of Science:

Biotechnology
Molecular Biology
Histology

Background:

Immunohistochemistry (IHC) is a sensitive technique for protein detection in biological tissues.
It relies on the specific binding of antibodies to target antigens.
IHC enables visualization of protein abundance, distribution, and localization within the cellular context (in situ).

Purpose of the Study:

To provide a comprehensive overview of standard immunohistochemistry protocols.
To detail the use of chromogenic substrates for protein detection in IHC.
To offer guidance on troubleshooting common issues encountered during IHC procedures.

Main Methods:

Discussion of pre-treatment steps for tissue samples.
Explanation of various types of chromogenic substrates used in IHC.
Identification and solutions for troubleshooting common IHC protocol stages.

Main Results:

Standardized protocols for effective IHC analysis.
Understanding of chromogenic substrate selection and application.
Improved troubleshooting capabilities for IHC experiments.

Conclusions:

Immunohistochemistry is a powerful tool for in situ protein analysis.
Adherence to optimized protocols and troubleshooting is crucial for reliable IHC results.
This chapter serves as a practical guide for researchers utilizing IHC with chromogenic detection.