4.4 Article

Collecting large datasets of rotational electron diffraction with ParallEM and SerialEM

Journal

JOURNAL OF STRUCTURAL BIOLOGY
Volume 211, Issue 2, Pages -

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.jsb.2020.107549

Keywords

Automated data collection; ParallEM; Electron diffraction; Electron 3D crystallography; CRYO ARM; 3D ED/MicroED

Funding

  1. Japan Society for the Promotion of Science [16H04757, 24657111]
  2. RIKEN Pioneering Project, Dynamic Structural Biology
  3. Cyclic Innovation for Clinical Empowerment (CiCLE) from the Japan Agency for Medical Research and Development, AMED
  4. JST CREST, Japan [JPMJCR18J2]
  5. Grants-in-Aid for Scientific Research [16H04757] Funding Source: KAKEN

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A semi-automated protocol has been developed for rotational data collection of electron diffraction patterns by combined use of SerialEM and ParallEM, where SerialEM is used for positioning of sample crystals and ParallEM for rotational data collection. ParallEM calls standard camera control software through an AutoIt script, which adapts to software operational changes and to new GUI programs guiding other cameras. Development included periodic flashing and pausing of data collection during overnight or day-long recording with a cold field-emission beam. The protocol proved to be efficient and accurate in data collection of large-scale rotational series from two JEOL electron microscopes, a general-purpose JEM-2100 and a high-end CRYO ARM 300. Efficiency resulted from simpler steps and task specialization. It is possible to collect 12-20 rotational series from similar to- 68 degrees to similar to 68 degrees at a rotation speed of 1 degrees/s in one hour without human supervision.

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