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强大的单颗粒冷冻电子显微镜 (cryo-EM) 处理工作流程,具有冷冻 SPARC、RELION 和 Scipion
JoVE Journal
Bioquímica
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JoVE Journal Bioquímica
A Robust Single-Particle Cryo-Electron Microscopy (cryo-EM) Processing Workflow with cryoSPARC, RELION, and Scipion
DOI:

13:43 min

January 31, 2022

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Capítulos

  • 00:04Introduction
  • 00:56Creating a New CryoSPARC v3 Project and Importing Data
  • 01:34CryoSPARC v3: Movie Alignment and Contrast Transfer Function (CTF) Estimation
  • 02:52CryoSPARC v3: Manual and Template-Based Particle Picking
  • 04:21CryoSPARC v3: 2D Classification
  • 04:52CryoSPARC v3: Ab-initio Reconstruction and Homogeneous Refinement
  • 05:31Exporting Particle Coordinates from CryoSPARC v3 and Importing Them to RELION-3 Using PyEM
  • 05:56RELION-3: Particle Extraction and 2D Classification, 3D Refinement, Mask Creation, and Post-Processing
  • 07:41RELION-3: Polishing Training, Particle Polishing, CTF, and Per-Particle Refinements
  • 09:15Transferring RELION-3 Particle Coordinates and 3D Map to Scipion 3
  • 10:05Scipion 3: High-Resolution Refinement
  • 11:21Results: Using cryoSPARC v3, RELION-3, and Scipion 3 to Obtain a High-Resolution 3D Reconstruction of the Adeno-Associated Virus
  • 12:52Conclusion

Summary

Tadução automática

本文介绍了如何有效利用三种冷冻电镜处理平台,即 cryoSPARC v3、RELION-3 和 Scipion 3,创建适用于各种单颗粒数据集的单一且强大的工作流程,以实现高分辨率结构测定。

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