Journal
/
/
cryoSPARC, RELION 및 Scipion을 사용한 견고한 단일 입자 Cryo-Electron Microscopy (cryo-EM) 처리 워크플로우
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
A Robust Single-Particle Cryo-Electron Microscopy (cryo-EM) Processing Workflow with cryoSPARC, RELION, and Scipion
DOI:

13:43 min

January 31, 2022

,

Chapters

  • 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

Automatic Translation

이 문서에서는 3개의 cryo-EM 처리 플랫폼(예: cryoSPARC v3, RELION-3 및 Scipion 3)을 효과적으로 활용하여 고해상도 구조 결정을 위한 다양한 단일 입자 데이터 세트에 적용할 수 있는 단일하고 강력한 워크플로우를 만드는 방법을 설명합니다.

Related Videos

Read Article