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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.
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.
The...
Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

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Related Experiment Video

Updated: Jun 4, 2026

The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells
11:45

The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells

Published on: February 27, 2020

Immunolabeling for electron microscopy.

C E Sarraf1

  • 1Department of Histology, Imperial College School of Medicine, London, UK.

Methods in Molecular Medicine
|February 22, 2011
PubMed
Summary
This summary is machine-generated.

Postembedding immunolabeling for transmission electron microscopy (TEM) requires careful antigenicity assessment using light microscopy first. If an antigen is undetectable by light microscopy, TEM methods will also fail.

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Last Updated: Jun 4, 2026

The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells
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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Immunohistochemistry

Background:

  • Immunolabeling identifies antigen location using specific antibodies.
  • Electron microscopy (EM) offers ultrastructural localization of antigens.
  • EM visualization requires electron-dense heavy metals, unlike light microscopy's colored products.

Purpose of the Study:

  • To outline protocols for postembedding immunolabeling in transmission electron microscopy (TEM).
  • To emphasize the importance of pre-validation of antigen presence using light microscopy.
  • To discuss challenges in maintaining antigenicity during harsh EM tissue processing.

Main Methods:

  • Postembedding immunolabeling protocols for TEM.
  • Light microscopy assessment of antigen presence in wax-embedded sections.
  • Comparison with pre-embedding and frozen tissue techniques.

Main Results:

  • Antigen presence must be confirmed by light microscopy before proceeding to EM.
  • Failure to detect an antigen with light microscopy precludes its detection with EM.
  • Tissue processing for EM is more demanding than for light microscopy, impacting antigenicity.

Conclusions:

  • Successful ultrastructural antigen localization via TEM relies on initial light microscopy validation.
  • Pre-embedding and frozen tissue methods offer alternatives when standard EM protocols fail.
  • Rigorous tissue preparation is crucial for preserving antigenicity in EM immunolabeling.