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Related Concept Videos

Immunogold Electron Microscopy01:20

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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Overview of Microscopy Techniques

The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...
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Immunofluorescence Microscopy01:12

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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Nanogold Labeling of the Yeast Endosomal System for Ultrastructural Analyses
09:49

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Immunogold probes in electron microscopy.

J E Beesley1

  • 1Wellcome Research Laboratories, Beckenham, UK.

Methods in Molecular Biology (Clifton, N.J.)
|November 15, 2012
PubMed
Summary
This summary is machine-generated.

Electron microscopy and immunocytochemistry reveal cell structures but require balancing preservation of cell details with antigen integrity. Optimizing sample preparation is key for accurate ultrastructural and antigen localization studies.

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Last Updated: May 16, 2026

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Immunocytochemistry

Background:

  • Electron microscopy (EM) enables high-resolution visualization of cellular ultrastructure and tissue architecture.
  • Electron immunocytochemistry (EIC) combines EM with antibody-based detection for precise localization of antigens within cells.
  • A fundamental challenge in EIC is the conflict between fixation methods optimal for ultrastructural preservation and those that maintain antigenicity.

Purpose of the Study:

  • To highlight the critical compromise in cell preparation for electron microscopy.
  • To discuss the balance between preserving ultrastructure and antigenicity for electron immunocytochemistry.

Main Methods:

  • Review of established protocols for electron microscopy sample preparation.
  • Analysis of the impact of fixation techniques on antigen integrity and ultrastructural detail.

Main Results:

  • Optimal fixation for EM fine structure often leads to antigen damage, reducing antibody binding.
  • Conversely, methods preserving antigenicity may compromise cellular ultrastructure.

Conclusions:

  • Cell preparation for EM and EIC necessitates a careful balance to achieve meaningful results.
  • Compromise is essential to retain sufficient antigenicity while preserving the necessary cell ultrastructure for high-resolution analysis.