Chapter 16
Automation of Continuous-Rotation Data Collection
for MicroED
M. Jason de la Cruz
Abstract
Automated coordination of microscope and camera functions for MicroED data collection simplifies the
procedure for robust dataset acquisition and enables unattended sequential collection of many crystal
targets. This chapter discusses the prerequisites for an algorithm of data collection automation for
continuous-rotation MicroED and presents a practical protocol for achieving this goal using the popular
TEM control software program SerialEM.
Key words MicroED, CryoEM, Electron microscopy, Microcrystal, Electron diffraction, 3D, Electron crystallography
1 Introduction
Data collection for continuous-rotation microcrystal electron diffraction (MicroED) involves (a) microscope and software setup;
(b) nanocrystal screening on transmission electron microscope
(TEM) grids (Fig. 1a, b); (c) testing crystals for diffraction
(Fig. 1c, d); (d) preparing the crystals for data collection; and
(e) coordination of the microscope’s beam blanker/column valves,
stage rotation, and camera recording for data acquisition. Automating these actions by computer requires these steps to occur in a
strictly linear fashion. An experienced microscopist/crystallographer may prefer to identify, prepare for, and collect each crystal
manually because crystals on the same grid could have different
sizes and available tilt range. For a regular user, automation of data
acquisition advantageously reduces the chances for operator error
during data collection and frees up the user’s own time, particularly
when multiple datasets are collected.
Several publicly available software packages exist for the automated collection of images for transmission electron microscopy
[1], but many have limited provisions for full microscope control in
diffraction mode. As of July 2019, the following software packages
Tamir Gonen and Brent L. Nannenga (eds.), CryoEM: Methods and Protocols, Methods in Molecular Biology, vol. 2215,
https://doi.org/10.1007/978-1-0716-0966-8_16, © Springer Science+Business Media, LLC, part of Springer Nature 2021
321
Automation of Continuous-Rotation Data Collection
for MicroED
M. Jason de la Cruz
Abstract
Automated coordination of microscope and camera functions for MicroED data collection simplifies the
procedure for robust dataset acquisition and enables unattended sequential collection of many crystal
targets. This chapter discusses the prerequisites for an algorithm of data collection automation for
continuous-rotation MicroED and presents a practical protocol for achieving this goal using the popular
TEM control software program SerialEM.
Key words MicroED, CryoEM, Electron microscopy, Microcrystal, Electron diffraction, 3D, Electron crystallography
1 Introduction
Data collection for continuous-rotation microcrystal electron diffraction (MicroED) involves (a) microscope and software setup;
(b) nanocrystal screening on transmission electron microscope
(TEM) grids (Fig. 1a, b); (c) testing crystals for diffraction
(Fig. 1c, d); (d) preparing the crystals for data collection; and
(e) coordination of the microscope’s beam blanker/column valves,
stage rotation, and camera recording for data acquisition. Automating these actions by computer requires these steps to occur in a
strictly linear fashion. An experienced microscopist/crystallographer may prefer to identify, prepare for, and collect each crystal
manually because crystals on the same grid could have different
sizes and available tilt range. For a regular user, automation of data
acquisition advantageously reduces the chances for operator error
during data collection and frees up the user’s own time, particularly
when multiple datasets are collected.
Several publicly available software packages exist for the automated collection of images for transmission electron microscopy
[1], but many have limited provisions for full microscope control in
diffraction mode. As of July 2019, the following software packages
Tamir Gonen and Brent L. Nannenga (eds.), CryoEM: Methods and Protocols, Methods in Molecular Biology, vol. 2215,
https://doi.org/10.1007/978-1-0716-0966-8_16, © Springer Science+Business Media, LLC, part of Springer Nature 2021
321