biostudies-literatureMcMullan GMedical Research CouncilWellcome TrustBiotechnology and Biological Sciences Research Councile2312905120https://www.ebi.ac.uk/biostudies/studies/S-EPMC10710074biostudies-literatureUnknown120(49)Electron cryomicroscopy can, in principle, determine the structures of most biological molecules but is currently limited by access, specimen preparation difficulties, and cost. We describe a purpose-built instrument operating at 100 keV-including advances in electron optics, detection, and processing-that makes structure determination fast and simple at a fraction of current costs. The instrument attains its theoretical performance limits, allowing atomic resolution imaging of gold test specimens and biological molecular structure determination in hours. We demonstrate its capabilities by determining the structures of eleven different specimens, ranging in size from 140 kDa to 2 MDa, using a fraction of the data normally required. CryoEM with a microscope designed specifically for high-efProceedings of the National Academy of Sciences of the United States of AmericaStructure determination by cryoEM at 100 keV.PMC10710074MC_UP_1201/17220526/B/20/ZMC_UP_A025_1012BB/T003677/1MC_U105184322MC_UP_1201/15Hutchings KACannone GSchulze-Briese CWells TRusso CJMeffert MWilson HPeet MJEl-Gomati MMHenderson RMihaylov DDickerson JLGittins ONaydenova KLee YChen SSobhy MAYamashita KMcMullan GDalby JfalseStructure determination by cryoEM at 100 keV.Electron cryomicroscopy can, in principle, determine the structures of most biological molecules but is currently limited by access, specimen preparation difficulties, and cost. We describe a purpose-built instrument operating at 100 keV-including advances in electron optics, detection, and processing-that makes structure determination fast and simple at a fraction of current costs. The instrument attains its theoretical performance limits, allowing atomic resolution imaging of gold test specimens and biological molecular structure determination in hours. We demonstrate its capabilities by determining the structures of eleven different specimens, ranging in size from 140 kDa to 2 MDa, using a fraction of the data normally required. CryoEM with a microscope designed specifically for high-ef2023-01-01T00:00:00Z2023 Dec2026-04-12T23:18:08.34Z2025-02-19T01:18:24.828ZS-EPMC107100743801157310.1073/pnas.2312905120