Efremov, R. G. & Stroobants, A. Coma-corrected rapid single-particle cryo-EM data collection on the CRYO ARM 300. Acta Crystallographica Section D: Structural Biology, 2021, 77
Single-particle cryogenic electron microscopy has recently become a major
method for determining the structures of proteins and protein complexes. This
has markedly increased the demand for throughput of high-resolution electron
microscopes, which are required to produce high-resolution images at high rates.
An increase in data-collection throughput can be achieved by using large beamimage
shifts combined with off-axis coma correction, enabling the acquisition
of multiple images from a large area of the EM grid without moving the
microscope stage. Here, the optical properties of the JEOL CRYO ARM 300
electron microscope equipped with a K3 camera were characterized under offaxis
illumination conditions. It is shown that efficient coma correction can be
achieved for beam-image shifts with an amplitude of at least 10 mm, enabling a
routine throughput for data collection of between 6000 and 9000 images per day.
Use of the benchmark for the rapid data-collection procedure (with beam-image
shifts of up to 7 mm) on apoferritin resulted in a reconstruction at a resolution of
1.7 A°. This demonstrates that the rapid automated acquisition of high-resolution
micrographs is possible using a CRYO ARM 300.