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Related Concept Videos

Preparation of Samples for Electron Microscopy01:20

Preparation of Samples for Electron Microscopy

7.6K
To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...
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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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Related Experiment Video

Updated: Mar 15, 2026

Sample Preparation by 3D-Correlative Focused Ion Beam Milling for High-Resolution Cryo-Electron Tomography
08:20

Sample Preparation by 3D-Correlative Focused Ion Beam Milling for High-Resolution Cryo-Electron Tomography

Published on: October 25, 2021

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Specimen Preparation for High-Resolution Cryo-EM.

L A Passmore1, C J Russo1

  • 1MRC Laboratory of Molecular Biology, Cambridge, United Kingdom.

Methods in Enzymology
|August 31, 2016
PubMed
Summary
This summary is machine-generated.

This study presents a systematic approach for preparing high-quality vitrified specimens for single-particle electron cryomicroscopy (cryo-EM). It details protocols and troubleshooting for obtaining optimal electron micrographs for biomolecular structure determination.

Keywords:
Cryo-EMElectron cryo-microscopyElectron tomographyProtein structureSingle-particle reconstructionSubstrates

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Cryo-electron Microscopy Specimen Preparation By Means Of a Focused Ion Beam
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Last Updated: Mar 15, 2026

Sample Preparation by 3D-Correlative Focused Ion Beam Milling for High-Resolution Cryo-Electron Tomography
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Biological Sample Preparation by High-pressure Freezing, Microwave-assisted Contrast Enhancement, and Minimal Resin Embedding for Volume Imaging
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Cryo-electron Microscopy Specimen Preparation By Means Of a Focused Ion Beam

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Area of Science:

  • Structural Biology
  • Biophysics
  • Microscopy Techniques

Background:

  • High-resolution imaging of biomolecules requires thin specimens stable in a transmission electron microscope vacuum.
  • Specimen preparation is a critical bottleneck in single-particle electron cryomicroscopy (cryo-EM) for structure determination.
  • Biological samples typically involve frozen aqueous solutions of the target complex.

Purpose of the Study:

  • To provide a systematic workflow for preparing vitrified specimens for cryo-EM.
  • To optimize the process from purified biomolecular complex to high-resolution electron micrographs.
  • To offer guidance on troubleshooting and avoiding common pitfalls in specimen preparation.

Main Methods:

  • Development of protocols for vitrifying biomolecular complexes in aqueous solution.
  • Testing various support materials, including all-gold and graphene grids.
  • Implementation of troubleshooting strategies for failed preparations.

Main Results:

  • A comprehensive guide for preparing vitrified specimens suitable for cryo-EM.
  • Techniques for achieving high-resolution electron micrographs.
  • Recommendations for utilizing current microscope, detector, and support technologies.

Conclusions:

  • Effective specimen preparation is crucial for successful cryo-EM structure determination.
  • This systematic approach enhances the quality of electron micrographs.
  • The study provides valuable resources for researchers in structural biology.