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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.

Cryo-electron Microscopy

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...

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Related Experiment Video

Updated: Jul 7, 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

Cryo-immunogold electron microscopy.

Peter J Peters¹, Erik Bos, Alexander Griekspoor

¹Netherlands Cancer Institute, Amsterdam, The Netherlands.

Current Protocols in Cell Biology

|January 30, 2008

Summary

This summary is machine-generated.

High-resolution cryo-immunogold electron microscopy offers sensitive detection of protein and antigen localization within cells. This method preserves antigenicity by avoiding harsh organic solvents, enabling ultrastructural topological biochemistry studies.

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Area of Science:

Cell Biology
Biochemistry
Microscopy

Background:

Subcellular localization of proteins is crucial for understanding cellular functions.
Existing immunodetection methods can compromise antigenicity due to harsh processing steps.

Purpose of the Study:

To describe a high-resolution cryo-immunogold electron microscopy technique for sensitive subcellular protein localization.
To enable topological biochemistry studies at the ultrastructural level.

Main Methods:

Chemical fixation of cells or tissues using aldehydes.
Embedding fixed material in gelatin.
Cryosectioning and mounting ultrathin sections on Formvar-coated grids.
High-resolution cryo-immunogold electron microscopy for immunodetection.

Main Results:

Achieved sensitive immunodetection of antigens on ultrathin sections.
Preserved protein antigenicity by omitting harsh organic solvents.
Enabled ultrastructural analysis of protein localization.

Conclusions:

Cryo-immunogold electron microscopy is a highly sensitive method for subcellular antigen localization.
The technique provides superior preservation of antigenicity compared to traditional methods.
Facilitates detailed ultrastructural studies of protein distribution and function.