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Cryo-Electron Tomography Remote Data Collection and Subtomogram Averaging
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Biologie
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Journal JoVE Biologie
Cryo-Electron Tomography Remote Data Collection and Subtomogram Averaging

Cryo-Electron Tomography Remote Data Collection and Subtomogram Averaging

DOI:
10.3791/63923-v

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08:55 min

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July 12, 2022

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, , ,
1Electron Bio-Imaging Centre,Diamond Light Source Ltd, Harwell Science & Innovation Campus, 2Division of Structural Biology, Wellcome Trust Centre for Human Genetics,University of Oxford, 3Chinese Academy of Medical Sciences Oxford Institute,University of Oxford

Chapitres

  • 00:04Introduction
  • 00:30Remote Cryo‐Electron Tomography (ET) Data Acquisition Using Tomo5
  • 04:31Cryo‐ET Subtomogram Averaging of Apoferritin Using emClarity
  • 07:45Results: High‐Resolution Cryo‐Electron Tomography Remote Data Acquisition
  • 08:27Conclusion

Summary

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Traduction automatique

The present protocol describes high-resolution cryo-electron tomography remote data acquisition using Tomo5 and subsequent data processing and subtomogram averaging using emClarity. Apoferritin is used as an example to illustrate detailed step-by-step processes to achieve a cryo-ET structure at 2.86 Å resolution.

Tags

Keywords: Cryo-electron TomographyRemote Data CollectionSubtomogram AveragingCROWETMicromoleculesNative Cellular EnvironmentTomo Five SoftwareImage Acquisition ParametersAtlasScreeningTarget SetupImage Shift CalibrationEucentric Height AdjustmentLow Dose Tomography Setup

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