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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.

Preparation of Samples for Electron Microscopy

Preparation of Samples for Electron Microscopy

To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...

Scanning Electron Microscopy

Scanning Electron Microscopy

A scanning electron microscope (SEM) is used to study the surface features of a sample by using an electron beam that scans the sample surface in a two-dimensional manner. Typically, areas between ~1 centimeter to 5 micrometers in width can be imaged. SEM can be used to image bacteria, viruses, tissues as well as larger samples like insects. Conventional SEM gives a magnification ranging from 20X to 30,000X and spatial resolution of 50 to 100 nanometers.
Fundamental Principles
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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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Related Experiment Video

Updated: Mar 16, 2026

Nanogold Labeling of the Yeast Endosomal System for Ultrastructural Analyses

Nanogold Labeling of the Yeast Endosomal System for Ultrastructural Analyses

Published on: July 14, 2014

Immunogold Labeling for Scanning Electron Microscopy.

Martin W Goldberg¹, Jindřiška Fišerová²

¹School of Biological and Biomedical Sciences, Durham University, South Road, Durham, DH1 3LE, UK. m.w.goldberg@durham.ac.uk.

Methods in Molecular Biology (Clifton, N.J.)

|August 13, 2016

Summary

This summary is machine-generated.

This study presents a generic method for immunogold labeling for scanning electron microscopy. This technique allows for high-resolution imaging and quantification of surface structures on various biological specimens.

Keywords:

Envelope Immunogold Labeling Nuclear Scanning electron microscopy Xenopus

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Last Updated: Mar 16, 2026

Nanogold Labeling of the Yeast Endosomal System for Ultrastructural Analyses

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Published on: July 14, 2014

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Post-embedding Immunogold Labeling of Synaptic Proteins in Hippocampal Slice Cultures

Post-embedding Immunogold Labeling of Synaptic Proteins in Hippocampal Slice Cultures

Published on: April 3, 2013

Area of Science:

Biological imaging
Microscopy techniques
Cell biology

Background:

Scanning electron microscopy (SEM) offers pseudo- or true 3D imaging of surfaces from whole organisms to macromolecules.
SEM magnifications range from 10× to 1,000,000×, enabling visualization of diverse biological structures.
Immunolabeling with antibodies can identify specific proteins and quantify their distribution in SEM.

Purpose of the Study:

To present a generic method for immunogold labeling for scanning electron microscopy (SEM).
To demonstrate the application of this method using isolated nuclear envelopes and mammalian cell cytoskeleton.
To discuss critical parameters for optimizing immunogold SEM results.

Main Methods:

Sample preparation and fixation protocols tailored for SEM.
Immunogold labeling using antibodies to target specific cellular components.
Post-labeling procedures including drying, metal coating, and SEM imaging.

Main Results:

Successful application of the generic immunogold labeling method to isolated nuclear envelopes.
Demonstration of visualizing the cytoskeleton of mammalian culture cells using immunogold SEM.
Identification and discussion of key parameters influencing image quality and data acquisition.

Conclusions:

The presented generic method is effective for immunogold labeling in SEM across different biological specimens.
Optimized sample preparation and labeling are crucial for high-resolution surface imaging.
Immunogold SEM provides a powerful approach for studying surface structures and protein localization in biology.