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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.

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Related Experiment Video

Updated: Jun 8, 2026

A high-throughput method to globally study the organelle morphology in S. cerevisiae

A high-throughput method to globally study the organelle morphology in S. cerevisiae

Published on: March 2, 2009

Correlative GFP-immunoelectron microscopy in yeast.

Christopher Buser¹, Kent McDonald

¹Department of Molecular and Cell Biology, University of California, Berkeley, California, USA.

Methods in Enzymology

|October 16, 2010

Summary

Correlative microscopy combines live-cell imaging with high-resolution ultrastructure. This study introduces a novel high-pressure freezing and freeze-substitution protocol for improved fine structure visibility and antigen preservation in correlative microscopy.

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Quantitative Live Cell Fluorescence-microscopy Analysis of Fission Yeast

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Last Updated: Jun 8, 2026

A high-throughput method to globally study the organelle morphology in S. cerevisiae

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Quantitative Live Cell Fluorescence-microscopy Analysis of Fission Yeast

Quantitative Live Cell Fluorescence-microscopy Analysis of Fission Yeast

Published on: January 23, 2012

Area of Science:

Cell Biology
Microscopy
Biochemistry

Background:

Correlative light and electron microscopy (CLEM) aims to integrate live-cell dynamics with ultrastructural details.
Existing CLEM techniques face challenges in preserving fine structures and antigenicity for immunolabeling.
Technical limitations hinder the practical application of combining fluorescence microscopy and immunoelectron microscopy.

Purpose of the Study:

To develop an improved protocol for correlative light and electron microscopy.
To achieve excellent fine structure visibility without compromising antigen integrity for immunolabeling.
To overcome technical limitations in current CLEM approaches.

Main Methods:

High-pressure freezing for rapid sample vitrification.
Fixative-free freeze-substitution to minimize structural damage and antigen disruption.
Low-temperature embedding for preserving antigenicity and fine structures.

Main Results:

The developed protocol enables excellent visibility of cellular fine structures.
Antigens recognized by immunolabeling protocols remain intact after the procedure.
The method addresses key challenges in combining live-cell imaging with high-resolution ultrastructural analysis.

Conclusions:

This high-pressure freezing and freeze-substitution protocol is a significant advancement for correlative microscopy.
It facilitates the visualization of cell biological processes by preserving both ultrastructure and antigenicity.
The protocol offers a more robust approach for integrating dynamic and structural information in cell biology research.