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Automation of Continuous-Rotation Data Collection for MicroED.

M Jason de la Cruz1

  • 1Structural Biology Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, NY, USA. delacrm1@mskcc.org.

Methods in Molecular Biology (Clifton, N.J.)
|December 28, 2020
PubMed
Summary

Automated data collection for Microcrystal Electron Diffraction (MicroED) simplifies robust dataset acquisition. This method enables unattended, sequential collection of multiple crystal targets using SerialEM software.

Keywords:
3DCryoEMElectron crystallographyElectron diffractionElectron microscopyMicroEDMicrocrystal

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Area of Science:

  • Crystallography
  • Electron Microscopy
  • Materials Science

Background:

  • Microcrystal Electron Diffraction (MicroED) is a powerful technique for determining the 3D structure of small crystals.
  • Manual data collection in MicroED can be time-consuming and requires significant user intervention.
  • Robust and efficient dataset acquisition is crucial for successful structure determination.

Purpose of the Study:

  • To present an automated data collection strategy for continuous-rotation MicroED.
  • To simplify the process of acquiring high-quality MicroED datasets.
  • To enable unattended, sequential data collection from multiple crystal targets.

Main Methods:

  • Development of an algorithm for automated data collection in MicroED.
  • Implementation of the automation protocol using SerialEM, a popular Transmission Electron Microscope (TEM) control software.
  • Focus on continuous-rotation MicroED data acquisition.

Main Results:

  • The automated coordination of microscope and camera functions simplifies MicroED data collection.
  • Robust dataset acquisition is achieved through automation.
  • Unattended, sequential collection of multiple crystal targets is enabled.

Conclusions:

  • Automated data collection protocols significantly enhance the efficiency and robustness of MicroED experiments.
  • The presented protocol using SerialEM provides a practical approach for automating MicroED data acquisition.
  • This advancement facilitates higher throughput structure determination using MicroED.