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À faire et à ne pas faire de cryo-microscopie électronique: Notions sur la préparation de l'échantillon et de collecte de données de haute qualité pour la reconstruction 3D macromoléculaire
JoVE Journal
Biologia
This content is Free Access.
JoVE Journal Biologia
Do’s and Don’ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction
DOI:

09:25 min

January 09, 2015

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Capítulos

  • 00:05Título
  • 01:21Cryo-holder Preparation and Maintenance
  • 02:32Grid Preparation
  • 03:48Plunge-freezing
  • 04:50Transfer the Cryo-grid to the TEM
  • 06:15Low-dose Data Collection
  • 07:03Results: Visualization of Macromolecules Using Cryo-electron Microscopy
  • 08:51Conclusion

Summary

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This paper describes the cryo-electron microscopy methodology, which is used to obtain high quality microscopic images of macromolecules in their near-native state. The method yields images suitable for further computerized processing using the single particle approach, devised to generate the 3D reconstruction of a macromolecule.

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