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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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Electron Microscope Tomography and Single-particle Reconstruction01:07

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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.
Electron Tomography
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Related Experiment Video

Updated: Aug 1, 2025

Single Particle Cryo-Electron Microscopy: From Sample to Structure
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Single Particle Cryo-Electron Microscopy: From Sample to Structure

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Protein Structural Analysis by Cryogenic Electron Microscopy.

Michael Hall1, Erin Schexnaydre2, Camilla Holmlund2

  • 1SciLifeLab Cryo-EM Facility, Department of Chemistry, Umeå University, Umeå, Sweden. michael.hall@umu.se.

Methods in Molecular Biology (Clifton, N.J.)
|April 24, 2023
PubMed
Summary
This summary is machine-generated.

This guide provides essential protocols for starting cryogenic electron microscopy (cryo-EM) projects. Learn the crucial first steps of specimen preparation and data collection for protein complex structure determination.

Keywords:
Cryo electron microscopyProtein structureSingle particle analysisVitrification

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Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction
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Area of Science:

  • Structural Biology
  • Biophysics

Background:

  • Cryogenic electron microscopy (cryo-EM) is a rapidly advancing technique for determining the structures of biomolecular assemblies.
  • Accurate structure determination is vital for understanding protein function and disease mechanisms.

Purpose of the Study:

  • To provide a detailed, step-by-step protocol for the initial stages of a cryo-EM project.
  • To guide researchers new to cryo-EM in specimen preparation and data collection.

Main Methods:

  • Detailed protocols for cryo-EM specimen preparation.
  • Step-by-step instructions for cryo-EM data collection.
  • A comprehensive list of required materials for cryo-EM experiments.

Main Results:

  • The study outlines a clear, actionable workflow for initiating cryo-EM studies.
  • Essential materials and procedural sequences are explicitly defined.

Conclusions:

  • These protocols serve as a foundational resource for researchers embarking on cryo-EM structure determination.
  • Successful implementation of these initial steps is critical for obtaining high-quality cryo-EM data.