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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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Immunohistochemistry.

Home
Immunohistochemistry.

Related Experiment Video

Histological-Based Stainings Using Free-Floating Tissue Sections

Histological-Based Stainings Using Free-Floating Tissue Sections

Published on: August 25, 2020

Immunohistochemistry.

Patricia A Loughran^1,2, Mark A Ross¹, Claudette M St Croix¹

¹University of Pittsburgh, Center for Biologic Imaging, Pittsburgh, Pennsylvania.

Current Protocols

|September 14, 2022

View abstract on PubMed

Summary

This summary is machine-generated.

This guide details immunohistochemistry methods for protein localization in cells and tissues. It covers techniques for adherent and suspension cells, tissue sections, and troubleshooting antibody detection.

Keywords:

antibodies and fluorophore imaging immunofluorescence microscopy

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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

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

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Published on: April 8, 2016

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

Area of Science:

Cell Biology
Biochemistry
Histology

Background:

Immunohistochemistry is crucial for protein analysis in biological samples.
Accurate protein localization and quantification are vital in research and diagnostics.

Purpose of the Study:

To provide comprehensive protocols for protein labeling using immunohistochemistry.
To guide researchers in selecting antibodies and detection methods.
To offer troubleshooting advice for common issues in immunohistochemistry.

Main Methods:

Detailed protocols for immunofluorescent labeling of adherent and suspension cells.
Methods for immunofluorescent labeling of tissue sections.
Techniques for double-labeling using various antibody combinations and streptavidin-biotin conjugates.

Main Results:

Established protocols enable effective protein localization in diverse biological samples.
Guidance on antibody selection and detection systems optimizes experimental outcomes.
Troubleshooting tips address common challenges, improving reproducibility.

Conclusions:

This article serves as a practical resource for performing immunohistochemistry.
The provided methods facilitate accurate protein analysis in cellular and tissue contexts.
Effective application of these techniques supports advancements in biological research.