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Related Concept Videos

Cryo-electron Microscopy01:28

Cryo-electron Microscopy

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Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...
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Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
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Related Experiment Video

Updated: May 4, 2026

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Cryo-electron microscopy of vitreous sections.

Petr Chlanda1, Martin Sachse

  • 1National Institute of Health, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 21, 2013
PubMed
Summary

Vitrification of pure water enables cryo-electron microscopy (cryo-EM) for near-native imaging. This guide details Cryo-Electron Microscopy of Vitreous Section (CEMOVIS) for imaging thicker biological samples using cryo-ultramicrotomy.

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Area of Science:

  • Biophysics
  • Microscopy techniques
  • Structural biology

Background:

  • Vitrification of pure water, previously thought impossible without cryoprotectants, revolutionized cryo-electron microscopy (cryo-EM).
  • Cryo-EM allows near-native observation of biological samples at the nanoscale.
  • Limitations in electron penetration restrict cryo-EM to thin samples, necessitating specialized techniques for larger biological specimens.

Purpose of the Study:

  • To provide a comprehensive, step-by-step protocol for Cryo-Electron Microscopy of Vitreous Section (CEMOVIS).
  • To enable imaging of thicker biological samples (cell or tissue level) using transmission electron microscopy (TEM).
  • To guide researchers in producing and imaging vitreous sections suitable for high-resolution cryo-EM analysis.

Main Methods:

  • Cryo-ultramicrotomy is employed to create thin sections (40-100 μm) of vitreous biological material.
  • Sample preparation involves trimming, sectioning with a diamond knife, and mounting onto an electron microscopy grid.
  • All procedures must be conducted below the devitrification temperature to maintain sample integrity.

Main Results:

  • Successful production of vitreous sections of biological samples suitable for cryo-EM.
  • Demonstration of CEMOVIS as a viable method for imaging bulky specimens.
  • Preservation of native structure in thicker biological samples through vitrification and cryo-sectioning.

Conclusions:

  • CEMOVIS is an effective technique for preparing thick biological samples for cryo-EM analysis.
  • The protocol ensures the preservation of sample integrity by maintaining cryogenic temperatures throughout the process.
  • This method expands the applicability of cryo-EM to a broader range of biological structures, including cells and tissues.