biostudies-literaturePeck JVNCI NIH HHSCore Facility Advocacy Committee and Office of Research Technologies, UNC Chapel Hill School of MedicineNational Cancer Institute of the National Institutes of Health243-252https://www.ebi.ac.uk/biostudies/studies/S-EPMC8895008biostudies-literatureUnknown9(Pt 2)Limitations to successful single-particle cryo-electron microscopy (cryo-EM) projects include stable sample generation, production of quality cryo-EM grids with randomly oriented particles embedded in thin vitreous ice and access to microscope time. To address the limitation of microscope time, methodologies to more efficiently collect data on a 200 keV Talos Arctica cryo-transmission electron microscope at speeds as fast as 720 movies per hour (∼17 000 per day) were tested. In this study, key parameters were explored to increase data collection speed including: (1) using the beam-image shift method to acquire multiple images per stage position, (2) employing UltrAufoil TEM grids with R0.6/1 hole spacing, (3) collecting hardware-binned data and (4) adjusting the image shift delay factor inIUCrJHigh-speed high-resolution data collection on a 200 keV cryo-TEM.PMC8895008P30CA016086P30 CA016086Strauss JDPeck JVFay JFfalseHigh-speed high-resolution data collection on a 200 keV cryo-TEM.Limitations to successful single-particle cryo-electron microscopy (cryo-EM) projects include stable sample generation, production of quality cryo-EM grids with randomly oriented particles embedded in thin vitreous ice and access to microscope time. To address the limitation of microscope time, methodologies to more efficiently collect data on a 200 keV Talos Arctica cryo-transmission electron microscope at speeds as fast as 720 movies per hour (∼17 000 per day) were tested. In this study, key parameters were explored to increase data collection speed including: (1) using the beam-image shift method to acquire multiple images per stage position, (2) employing UltrAufoil TEM grids with R0.6/1 hole spacing, (3) collecting hardware-binned data and (4) adjusting the image shift delay factor in2022-01-01T00:00:00Z2022 Mar2026-05-09T04:16:54.23Z2025-02-19T00:56:24.35ZS-EPMC88950083537150410.1107/S2052252522000069