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冷冻电子断层扫描远程数据收集和断层扫描平均
JoVE Journal
Biology
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JoVE Journal Biology
Cryo-Electron Tomography Remote Data Collection and Subtomogram Averaging
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冷冻电子断层扫描远程数据收集和断层扫描平均

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

Chapters

  • 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

Automatic Translation

Automatic Translation

本协议描述了使用Tomo5的高分辨率冷冻电子断层扫描远程数据采集以及使用emClarity的后续数据处理和子断层图平均。以载铁蛋白为例，说明实现 2.86 Å 分辨率冷冻电子断层扫描结构的详细分步过程。

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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