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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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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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Immunoprecipitation

Immunoprecipitation

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Chromatin Immunoprecipitation
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Related Experiment Video

Updated: Jun 4, 2026

The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells

The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells

Published on: February 27, 2020

Ultrastructural immunochemistry.

Jeremy N Skepper, Janet M Powell

|March 2, 2011

Summary

This summary is machine-generated.

Colloidal gold technology revolutionized immunochemistry, enabling precise visualization of antigens using transmission electron microscopy (TEM). This guide details essential considerations for successful immunogold staining, from antigen assessment to fixation and controls.

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Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography

Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography

Published on: March 9, 2010

Area of Science:

Immunochemistry
Electron Microscopy
Biotechnology

Background:

Colloidal gold technology is a cornerstone in immunochemistry development.
Gold particles offer strong electron scattering for clear visualization in transmission electron microscopy (TEM).
Successful immunogold staining requires thorough understanding of antibody-antigen characteristics and localization.

Purpose of the Study:

To outline critical methods and considerations for effective immunogold staining.
To guide researchers in optimizing immunogold staining protocols for antigen detection.

Main Methods:

Assessing antigen location (extracellular, intracellular, membrane-associated, soluble).
Evaluating antigen quantity and subcellular compartment sequestration.
Considering antigen vulnerability to fixation and embedding procedures.

Main Results:

Antibody specificity on Western blots does not guarantee immunochemistry utility.
Higher antibody concentrations are often needed for immunofluorescence and immunogold staining compared to Western blots.
Some antibodies are unsuitable for immunochemistry applications.

Conclusions:

Immunogold staining requires careful planning regarding antigen properties and antibody behavior.
Key factors for successful application include proper fixation, appropriate controls, and optimized resolution and quantification strategies.