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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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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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On the Development of Electron Cryo-Microscopy (Nobel Lecture).

Jacques Dubochet1

  • 1UNIL - Université de Lausanne, Fac. of Biology and Medicine, Deptm. of Ecology and Evolution, 1015, Lausanne, Switzerland.

Angewandte Chemie (International Ed. in English)
|July 10, 2018
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Summary

Electron microscopy requires samples to be dry, causing molecules to stick. Vitrifying water suspension enables electron-cryo microscopy by preserving biological specimens.

Keywords:
Nobel lectureelectron cryo-microscopyprotein structuresvitrification

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Area of Science:

  • Biophysics
  • Microscopy
  • Structural Biology

Background:

  • Electron microscopes require a vacuum environment, necessitating dry biological specimens.
  • Drying biological samples causes molecular adhesion, hindering high-resolution imaging.
  • Overcoming this limitation is crucial for visualizing cellular structures.

Purpose of the Study:

  • To present the solution enabling electron-cryo microscopy.
  • To explain the principle of preserving biological specimens for electron microscopy.

Main Methods:

  • Specimen suspension in vitrifying water.
  • Rapid cooling to prevent ice crystal formation.

Main Results:

  • Vitrifying water prevents ice crystal formation during rapid cooling.
  • Preserved molecular structure of biological specimens in an hydrated state.
  • Enabled high-resolution imaging of biological samples via electron microscopy.

Conclusions:

  • Suspension in vitrifying water is key to electron-cryo microscopy.
  • This method overcomes the drying artifact in electron microscopy.
  • Facilitates detailed structural analysis of biological molecules.