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Related Concept Videos

Confocal Fluorescence Microscopy01:16

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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Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
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Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions
13:43

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Published on: June 24, 2013

High data output method for 3-D correlative light-electron microscopy using ultrathin cryosections.

Katia Cortese1, Giuseppe Vicidomini, Maria Cristina Gagliani

  • 1MicroscoBio Research Center, Department of Experimental Medicine (DIMES), University of Genoa, Genoa, Italy.

Methods in Molecular Biology (Clifton, N.J.)
|October 23, 2012
PubMed
Summary

High Data Output CLEM (HDO-CLEM) combines fluorescence light microscopy and electron microscopy for detailed cellular analysis. This method enhances the correlation of molecular topography and ultrastructural morphology for high-throughput 3D imaging.

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Last Updated: May 17, 2026

Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions
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Correlative Confocal and 3D Electron Microscopy of a Specific Sensory Cell
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Correlative Confocal and 3D Electron Microscopy of a Specific Sensory Cell

Published on: July 19, 2015

Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Molecular Imaging

Background:

  • Investigating intracellular dynamics requires understanding molecular topography and cellular ultrastructure.
  • Standard fluorescence light microscopy (FLM) offers protein localization but lacks detailed cellular context.
  • Electron microscopy (EM) provides high resolution and cellular context but has limitations in statistical output and applicability.

Purpose of the Study:

  • To describe the High Data Output CLEM (HDO-CLEM) method for bridging FLM and EM.
  • To enable high-throughput correlation of molecular and ultrastructural data.
  • To facilitate hybrid morphometric analysis combining FLM and EM capabilities.

Main Methods:

  • Development and optimization of the HDO-CLEM approach.
  • Utilizing EM cryo-immunogold for extended analysis.
  • Implementing 3D FLM reconstruction software for semi/fully automatic analysis.

Main Results:

  • HDO-CLEM allows simultaneous correlation of hundreds of events.
  • The method supports three-dimensional (3D) correlation and immunolabeling of both endogenous and tagged proteins.
  • Optimized sample preparation and analysis routines were established.

Conclusions:

  • HDO-CLEM significantly enhances correlative light-electron microscopy capabilities.
  • This method combines the strengths of FLM and EM for precise, high-data-output cellular analysis.
  • It paves the way for advanced hybrid light/electron microscopy morphometry.