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The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.
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Related Experiment Video

Updated: Jun 27, 2025

Dual-color Correlative Light and Electron Microscopy for the Visualization of Interactions between Mitochondria and Lysosomes
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Some Guiding Principles for a "Simple" Correlative Light Electron Microscopy Experiment.

Elina Mäntylä1, Paul Verkade2

  • 1BioMediTech, Faculty of Medicine and Health Technology, Tampere University, Tampere, Finland.

Methods in Molecular Biology (Clifton, N.J.)
|May 6, 2024
PubMed
Summary
This summary is machine-generated.

Correlative Multimodal Imaging (CMI), especially correlative light and electron microscopy (CLEM), combines techniques for enhanced sample analysis. This study details a straightforward CLEM workflow for visualizing fluorescent proteins within cells.

Keywords:
CLEMEM correlationFluorescence microscopyImmunogold labellingMultimodal imagingSample preparation

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Area of Science:

  • Life Sciences
  • Microscopy
  • Cell Biology

Background:

  • Correlative Multimodal Imaging (CMI) integrates data from diverse imaging techniques to reveal insights beyond single-modality analysis.
  • Correlative Light and Electron Microscopy (CLEM) is a prominent CMI technique, merging light microscopy (LM) and transmission electron microscopy (TEM).
  • CLEM enables the precise localization of specific cellular components, such as fluorescently tagged proteins, with nanometer resolution.

Purpose of the Study:

  • To outline a simplified workflow for performing correlative light and electron microscopy (CLEM).
  • To describe the essential instrumentation and fundamental principles of sample preparation for CLEM experiments.
  • To demonstrate a practical CLEM approach utilizing stable expression of fluorescent proteins.

Main Methods:

  • Detailed description of the correlative light and electron microscopy (CLEM) workflow.
  • Explanation of instrumentation required for both light and electron microscopy stages.
  • Methodology for sample preparation, focusing on stable expression of fluorescent proteins for CLEM.

Main Results:

  • Successful implementation of a straightforward CLEM protocol.
  • Demonstration of correlative imaging capabilities for visualizing fluorescently labeled structures.
  • Achieved nanometer-resolution localization of intracellular components via CLEM.

Conclusions:

  • The described CLEM approach provides a feasible method for advanced cellular imaging.
  • This workflow facilitates the detailed study of protein localization and cellular ultrastructure.
  • Correlative Multimodal Imaging, specifically CLEM, offers significant advantages for biological research.