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Related Concept Videos

Cryo-electron Microscopy01:28

Cryo-electron Microscopy

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...
Transmission Electron Microscopy01:15

Transmission Electron Microscopy

In 1931, physicist Ernst Ruska—building on the idea that magnetic fields can direct an electron beam just as lenses can direct a beam of light in an optical microscope—developed the first prototype of the electron microscope. This development led to the development of the field of electron microscopy. In the transmission electron microscope (TEM), electrons are produced by a hot tungsten element and accelerated by a potential difference in an electron gun, which gives them up to 400 keV in...
Preparation of Samples for Electron Microscopy01:20

Preparation of Samples for Electron Microscopy

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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Related Experiment Video

Updated: Jun 8, 2026

Miniaturized Sample Preparation for Transmission Electron Microscopy
09:04

Miniaturized Sample Preparation for Transmission Electron Microscopy

Published on: July 27, 2018

Radiation damage in electron cryomicroscopy.

Lindsay A Baker1, John L Rubinstein

  • 1Molecular Structure and Function Program, The Hospital for Sick Children, Ontario, Canada.

Methods in Enzymology
|October 5, 2010
PubMed
Summary
This summary is machine-generated.

Electron beam radiation damages biological specimens during imaging, limiting resolution in cryo-electron microscopy (cryo-EM). Minimizing electron exposure and cooling samples reduces this radiation damage, optimizing image quality.

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Area of Science:

  • Structural Biology
  • Microscopy Techniques
  • Radiation Physics

Background:

  • Electron microscopy (EM) is crucial for visualizing biological structures at high resolution.
  • The electron beam used in EM causes radiation damage to biological specimens, limiting achievable resolution.
  • This damage is a fundamental bottleneck in biological cryo-electron microscopy (cryo-EM).

Purpose of the Study:

  • To review the mechanisms of radiation damage in cryo-EM.
  • To discuss practical strategies for minimizing radiation damage to biological specimens.
  • To provide guidance on optimizing imaging parameters for better resolution.

Main Methods:

  • Review of existing literature on electron-specimen interactions and radiation damage.
  • Analysis of the three stages of radiation damage: primary, secondary, and tertiary.
  • Discussion of mitigation strategies including dose reduction and temperature control.

Main Results:

  • Radiation damage occurs through ionization, bond breakage, secondary electron effects, and gas evolution.
  • Damage mechanisms are influenced by electron energy, specimen composition, and temperature.
  • Effective strategies involve limiting electron exposure and maintaining low specimen temperatures.

Conclusions:

  • Radiation damage is an inherent limitation in cryo-EM, affecting image resolution.
  • Careful management of electron exposure and specimen temperature is critical for high-resolution imaging.
  • Optimizing imaging parameters can significantly mitigate radiation damage effects.