TY - JOUR
T1 - Collecting large datasets of rotational electron diffraction with ParallEM and SerialEM
AU - Takaba, Kiyofumi
AU - Maki-Yonekura, Saori
AU - Yonekura, Koji
N1 - Funding Information:
We thank Tasuku Hamaguchi for feedback on use of ParallEM, David Mastronarde for technical assistance with SerialEM, Radostin Danev for SerialEM setup in single particle data collection, Keitaro Yamashita for usage of KAMO, Satoru Inoue and Tatsuo Hasegawa for providing crystal samples of organic molecules. This work was partly supported by Japan Society for the Promotion of Science Grant-in-Aid for Scientific Research Grant 16H04757 (to K.Y.), Japan Society for the Promotion of Science Grant-in-Aid for Challenging Exploratory Research Grant 24657111 (to K.Y.), the RIKEN Pioneering Project, Dynamic Structural Biology (to K.T., S.M.-Y. and K.Y.), the Cyclic Innovation for Clinical Empowerment (CiCLE) from the Japan Agency for Medical Research and Development, AMED (to K.Y.), and JST CREST Grant Number JPMJCR18J2, Japan (to K.T., S. M.-Y., and K. Y.).
Publisher Copyright:
© 2020 Elsevier Inc.
PY - 2020/8/1
Y1 - 2020/8/1
N2 - 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 ~−68° to ~68° at a rotation speed of 1°/s in one hour without human supervision.
AB - 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 ~−68° to ~68° at a rotation speed of 1°/s in one hour without human supervision.
KW - 3D ED/MicroED
KW - Automated data collection
KW - CRYO ARM
KW - Electron 3D crystallography
KW - Electron diffraction
KW - ParallEM
UR - http://www.scopus.com/inward/record.url?scp=85086586862&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85086586862&partnerID=8YFLogxK
U2 - 10.1016/j.jsb.2020.107549
DO - 10.1016/j.jsb.2020.107549
M3 - Article
C2 - 32544623
AN - SCOPUS:85086586862
SN - 1047-8477
VL - 211
JO - Journal of Structural Biology
JF - Journal of Structural Biology
IS - 2
M1 - 107549
ER -