Date Published: April 01, 2020
Publisher: International Union of Crystallography
Author(s): Kasim Sader, Rishi Matadeen, Pablo Castro Hartmann, Tor Halsan, Chris Schlichten.
The setup and operation of an industrial cryo-EM laboratory is described.
Transmission electron microscopy (TEM) laboratory designs and cryo-EM facility management play a crucial role in the optimal operation of the microscopes and their output (O’Keefe et al., 2004 ▸; Muller et al., 2006 ▸; Martínez, 2014 ▸; Alewijnse et al., 2017 ▸; Clare et al., 2017 ▸; Kandiah et al., 2019 ▸). However, laboratory-design reports are focused largely on materials science and facility management on academic cryo-EM. The present report aims to address laboratory design and facility management both specifically for single-particle cryo-EM and in an industrial context.
In terms of facility operation, vitrification with the Vitrobot Mk IV is very reproducible. Krios alignments change very little over time with stable temperature, and the only routine alignments that are performed are beam shift, objective aperture centering, and automated coma and astigmatism correction. A significant investment in IT expertise is required to store and transfer the large volumes of data produced and to facilitate timely and efficient image processing. Some on-the-fly pre-processing is performed, which is useful for monitoring changes in instrument performance and quickly starting 2D analysis or 3D refinement. With changes to cryo-cycling procedures, we achieved the use of 355 out of 365 days for the Consortium Krios 1 in 2018. These points are generally applicable to both industrial and academic facilities.