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Related Concept Videos

Immunocytochemistry and Immunohistochemistry01:22

Immunocytochemistry and Immunohistochemistry

11.6K
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...
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Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

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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.
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Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

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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: May 3, 2026

Histological-Based Stainings Using Free-Floating Tissue Sections
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Histological-Based Stainings Using Free-Floating Tissue Sections

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Immunohistochemistry.

Florence M Hofman1, Clive R Taylor1

  • 1University of Southern California, Los Angeles, California.

Current Protocols in Immunology
|February 11, 2014
PubMed
Summary
This summary is machine-generated.

This guide details immunohistochemistry (IHC) methods for antigen localization in tissues and cells. It covers sample preparation, multi-antigen detection, and troubleshooting techniques like antigen retrieval and preabsorption for accurate results.

Keywords:
antigen retrievaldouble stainingimmunochemistryimmunohistochemistrystaining cell culturesstaining fixed tissuesstaining frozen tissues

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Histology

Background:

  • Immunohistochemistry (IHC) is a powerful technique for visualizing antigen localization within biological samples.
  • Standard IHC protocols require careful optimization for different tissue types and antigen sensitivities.

Purpose of the Study:

  • To describe various methods for antigen localization using immunohistochemistry (IHC).
  • To provide guidance on sample preparation, multi-antigen detection, and troubleshooting common IHC issues.

Main Methods:

  • Detailed protocols for preparing fresh, frozen, and fixed paraffin-embedded tissues and cells.
  • Strategies for simultaneous detection of multiple antigens.
  • Methods for antigen retrieval and secondary antibody preabsorption to enhance specificity and sensitivity.

Main Results:

  • Successful localization of specific antigens in diverse biological preparations.
  • Demonstration of simultaneous detection of two or three antigens.
  • Identification of causes for background staining and methods for its elimination.

Conclusions:

  • Multiple IHC protocols are available, each with specific advantages and disadvantages.
  • Careful selection of reagents and techniques, including antigen retrieval and preabsorption, is crucial for successful IHC.
  • Optimization of IHC methods ensures accurate antigen localization and minimizes background noise.