MicroED data collection with SerialEM

被引:44
作者
de la Cruz, M. Jason [1 ,2 ]
Martynowycz, Michael W. [1 ,3 ,4 ,5 ]
Hattne, Johan [1 ,3 ,4 ,5 ]
Gonen, Tamir [1 ,3 ,4 ,5 ]
机构
[1] Howard Hughes Med Inst, Janelia Res Campus,19700 Helix Dr, Ashburn, VA 20147 USA
[2] Mem Sloan Kettering Canc Ctr, Struct Biol Program, Sloan Kettering Inst, 1275 York Ave, New York, NY 10065 USA
[3] Univ Calif Los Angeles, David Geffen Sch Med, Howard Hughes Med Inst, Los Angeles, CA 90095 USA
[4] Univ Calif Los Angeles, David Geffen Sch Med, Dept Biol Chem, Los Angeles, CA 90095 USA
[5] Univ Calif Los Angeles, David Geffen Sch Med, Dept Physiol, Los Angeles, CA 90095 USA
关键词
MicroED; Microcrystal; Electron diffraction; cryoEM; Transmission electron microscopy; SerialEM; DIFFRACTION DATA;
D O I
10.1016/j.ultramic.2019.03.009
中图分类号
TH742 [显微镜];
学科分类号
摘要
The cryoEM method Microcrystal Electron Diffraction (MicroED) involves transmission electron microscope (TEM) and electron detector working in synchrony to collect electron diffraction data by continuous rotation. We previously reported several protein, peptide, and small molecule structures by MicroED using manual control of the microscope and detector to collect data. Here we present a procedure to automate this process using a script developed for the popular open-source software package SerialEM. With this approach, SerialEM coordinates stage rotation, microscope operation, and camera functions for automated continuous-rotation MicroED data collection. Depending on crystal and substrate geometry, more than 300 datasets can be collected overnight in this way, facilitating high-throughput MicroED data collection for large-scale data analyses.
引用
收藏
页码:77 / 80
页数:4
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