Cryo‐EM—the first thirty years

@article{Dubochet2012CryoEMtheFT,
  title={Cryo‐EM—the first thirty years},
  author={Jacques Dubochet},
  journal={Journal of Microscopy},
  year={2012},
  volume={245}
}
  • J. Dubochet
  • Published 1 March 2012
  • Biology
  • Journal of Microscopy
Thirty years ago, in December 1981, The Journal of Microscopy published a very short paper entitled ‘Vitrification of pure water for electron microscopy’. It turned out to be important for the development of cryo‐electron microscopy and it contributed to reverse, from foe to friend, the status of water in electron microscopists’ minds. This change has brought obvious gains. The future will tell how many more are still to come. 

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This introductory chapter takes the perspective of the workflow of a cryo-tomography project to outline the technique, and important underlying concepts.

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Toward High-Resolution Cryo-Electron Microscopy: Technical Review on Microcrystal-Electron Diffraction

Cryo-electron microscopy (cryo-EM) is arguably the most powerful tool used in structural biology. It is an important analytical technique that is used for gaining insight into the functional and
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References

SHOWING 1-10 OF 47 REFERENCES

Cryo-electron microscopy of vitrified specimens.

Cryo-electron microscopy of vitrified specimens was just emerging as a practical method when Richard Henderson proposed that an EMBO course on the new technique was taught, and water, which was once the arch enemy of all electronmicroscopists, became what it always was in nature – an integral part of biological matter and a beautiful substance.

Cryo-electron microscopy of viruses

Cryo-electron microscopy of vitrified specimens offers possibilities for high resolution observations that compare favourably with any other electron microscopical method.

Cryo-electron microscopy of vitreous sections.

This chapter provides a step-by-step guide to produce and image vitreous sections of a biological sample using cryo-electron microscopy to observe biological samples at nanometer scale.

Reaching the information limit in cryo-EM of biological macromolecules: experimental aspects.

Golgi apparatus studied in vitreous sections

This work provides a first step towards the high‐resolution description of the secretory pathway in native vitrified samples and describes the challenges associated with this attempt.

Three-Dimensional Electron Microscopy of Macromolecular Assemblies: Visualization of Biological Molecules in Their Native State

Cryoelectron microscopy of biological molecules is among the hottest growth areas in biophysics and structural biology at present, and Frank is arguably the most distinguished practitioner of this

Analysis of cryo-electron microscopy images does not support the existence of 30-nm chromatin fibers in mitotic chromosomes in situ

It is strongly suggested that, within the bulk of compact metaphase chromosomes, the nucleosomal fiber does not undergo 30-nm folding, but exists in a highly disordered and interdigitated state, which is, on the local scale, comparable with a polymer melt.

Freezing, sectioning and observation artefacts of frozen hydrated sections for electron microscopy

I N T R O D U C T I O N In a recent article we have shown that thin sections of vitrified biological samples can be prepared, transferred and observed under good conditions in the electron microscope