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Related Concept Videos

Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

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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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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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Imaging Replicative Domains in Ultrastructurally Preserved Chromatin by Electron Tomography
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DNA Labeling at Electron Microscopy.

Nicolas Thelen1, Marc Thiry2

  • 1Giga Neurosciences, Department of Life Sciences, Laboratory of Cell Biology, University of Liège, Quartier Hôpital, 15 Avenue Hippocrate (B36), 4000, Liege, Belgium.

Methods in Molecular Biology (Clifton, N.J.)
|February 4, 2017
PubMed
Summary

This study introduces a new method for precisely locating DNA on ultrathin sections using terminal deoxynucleotidyl transferase (TdT) and non-isotopic nucleotide analogs, visualized with immunogold labeling.

Keywords:
DNA labelingImmuno-electron microscopyImmunocytochemistryImmunogold techniquesTerminal deoxynucleotidyl transferase

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Accurate DNA localization is crucial for understanding cellular processes.
  • Existing methods may have limitations in resolution or compatibility.

Purpose of the Study:

  • To develop a high-resolution method for pinpointing DNA location on ultrathin sections.
  • To create a technique compatible with standard biological sample preparation.

Main Methods:

  • Incubation of ultrathin sections with terminal deoxynucleotidyl transferase (TdT) and non-isotopic nucleotide analogs.
  • Visualization of labeled nucleotides via indirect immunogold labeling.
  • Compatibility with standard fixation and embedding procedures.

Main Results:

  • Successful high-resolution localization of DNA on ultrathin sections.
  • Demonstrated compatibility with various fixation and embedding techniques.
  • Effective visualization even in samples with low DNA amounts.

Conclusions:

  • The described method offers a powerful tool for precise DNA localization.
  • This technique enhances the ability to study DNA distribution in biological materials.
  • It is a versatile method that can be combined with cytochemical approaches.