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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.

Confocal Fluorescence Microscopy

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...

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Updated: May 4, 2026

Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles

Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles

Published on: August 7, 2016

Correlative light and electron microscopy using immunolabeled sections.

Heinz Schwarz¹, Bruno M Humbel

¹Max Planck Institute for Developmental Biology, Tübingen, Germany.

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

|December 21, 2013

Summary

This summary is machine-generated.

This chapter details electron microscopy preparation methods for optimal cellular ultrastructure preservation. These methods enable precise localization of macromolecules using correlative light and electron microscopy techniques.

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Area of Science:

Cell Biology
Microscopy Techniques
Biochemistry

Background:

Correlative microscopy integrates light and electron microscopy for cellular and subcellular analysis.
Standard preparation methods may not optimally preserve ultrastructure for high-resolution imaging.

Purpose of the Study:

To provide detailed, high-quality electron microscopy preparation protocols.
To ensure optimal preservation of cellular ultrastructure for correlative studies.
To facilitate accurate macromolecule localization within cells.

Main Methods:

Detailed electron microscopy sample preparation techniques.
Serial thin sectioning for comparative analysis.
Histochemical, immunofluorescence, and immunogold staining procedures.

Main Results:

Optimized preservation of cellular ultrastructure achieved.
Successful integration of light microscopy for orientation and region identification.
High-resolution localization of immunogold labels within cellular architecture.

Conclusions:

The presented electron microscopy preparation methods enhance ultrastructural preservation.
Correlative microscopy, utilizing these methods, allows precise macromolecule localization.
This approach is crucial for understanding cellular organization and molecular function.