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Related Concept Videos

Cryo-electron Microscopy01:28

Cryo-electron Microscopy

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Related Experiment Video

Updated: May 19, 2026

Correlative Light Electron Microscopy (CLEM) for Tracking and Imaging Viral Protein Associated Structures in Cryo-immobilized Cells
09:18

Correlative Light Electron Microscopy (CLEM) for Tracking and Imaging Viral Protein Associated Structures in Cryo-immobilized Cells

Published on: September 7, 2018

3D HDO-CLEM: cellular compartment analysis by correlative light-electron microscopy on cryosection.

Katia Cortese1, Giuseppe Vicidomini, Maria Cristina Gagliani

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

Methods in Cell Biology
|August 4, 2012
PubMed
Summary

High-data-output correlative light-electron microscopy (CLEM) combines fluorescence light microscopy and electron microscopy for detailed cell structure analysis. This method enables high-resolution imaging and large-scale data correlation for a comprehensive understanding of cellular processes.

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Biophysics

Background:

  • Fluorescence light microscopy (FLM) offers large field of view but limited resolution and cannot visualize unlabeled cellular context.
  • Electron microscopy (EM) provides high resolution and cellular context but has a limited field of view.
  • Correlative light-electron microscopy (CLEM) integrates FLM and EM to overcome individual limitations.

Purpose of the Study:

  • To describe a protocol for high-data-output CLEM (HDO-CLEM) using Tokuyasu cryosections.
  • To enable correlation of molecular topography with ultrastructural morphology at high resolution.
  • To combine the strengths of FLM and EM for comprehensive cellular analysis.

Main Methods:

  • Immunolabeling of Tokuyasu cryosections for correlative LM and EM imaging.
  • Optimization of critical sample preparation steps for ultrastructural preservation.
  • Development of software for semi/fully automatic 3D FLM reconstruction and hybrid morphometry.

Main Results:

  • HDO-CLEM allows correlation of hundreds of events simultaneously.
  • The method supports three-dimensional (3D) correlation and simultaneous immunolabeling of multiple protein types.
  • Integration of FLM's data analysis with EM's precision for hybrid morphometric analysis.

Conclusions:

  • HDO-CLEM provides a powerful approach for detailed cellular ultrastructure and molecular localization studies.
  • This method enhances the understanding of protein function relative to cell compartments and morphology.
  • The developed protocol and software facilitate advanced hybrid light/EM morphometry.