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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
Accelerated...

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.
The...

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Related Experiment Video

Updated: Jun 11, 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 labelling for scanning electron microscopy.

Martin W Goldberg¹, Jindriska Fiserova

¹School of Biological and Biomedical Sciences, Durham University, Durham, UK.

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

|July 6, 2010

Summary

This summary is machine-generated.

This study presents a method for immunogold labeling for scanning electron microscopy (SEM). This technique allows visualization of cellular components and macromolecules on various biological surfaces.

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Area of Science:

Biotechnology
Microscopy
Cell Biology

Background:

Scanning electron microscopy (SEM) offers high-resolution 3D imaging of biological surfaces, from whole organisms to macromolecules.
Labeling samples with antibodies enables identification and quantification of specific proteins at various magnifications.

Purpose of the Study:

To present a generic method for immunogold labeling specifically for scanning electron microscopy.
To demonstrate the application of this method on isolated nuclear envelopes and mammalian cell cytoskeletons.

Main Methods:

A generic protocol for immunogold labeling suitable for SEM was developed.
Sample preparation, fixation, antibody labeling, drying, metal coating, and imaging parameters were optimized.
The method was validated using isolated nuclear envelopes and mammalian cell cytoskeletons.

Main Results:

The study provides a detailed, adaptable protocol for immunogold SEM.
Successful visualization of surface structures and protein localization was achieved on the tested specimens.
Optimization of various parameters ensures high-quality results for diverse sample types.

Conclusions:

The presented immunogold labeling method is effective for high-resolution SEM imaging of biological samples.
This technique enhances the study of surface-related structures and protein distributions in cell biology.
The generic nature of the method allows for broad applicability across different biological specimens.