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TitleHigh-resolution cryo-EM using a common LaB 120-keV electron microscope equipped with a sub-200-keV direct electron detector.
Journal, issue, pagesSci Adv, Vol. 11, Issue 1, Page eadr0438, Year 2025
Publish dateJan 3, 2025
AuthorsHariprasad Venugopal / Jesse Mobbs / Cyntia Taveneau / Daniel R Fox / Ziva Vuckovic / Sahil Gulati / Gavin Knott / Rhys Grinter / David Thal / Stephen Mick / Cory Czarnik / Georg Ramm /
PubMed AbstractHigh-resolution cryo-electron microscopy (cryo-EM) requires costly 200- to 300-keV cryo-transmission electron microscopes (cryo-TEMs) with field emission gun (FEG) sources, stable columns, constant- ...High-resolution cryo-electron microscopy (cryo-EM) requires costly 200- to 300-keV cryo-transmission electron microscopes (cryo-TEMs) with field emission gun (FEG) sources, stable columns, constant-powered lenses, autoloader, and direct electron detectors (DED). Recent advances in 100-keV imaging with the emergence of sub-200-keV optimized DED technology promises the development of more affordable cryo-TEMs. So far, 100-keV imaging has required microscopes with FEG sources. We here explored whether a standard 120-keV TEMs with thermionic lanthanum hexaboride (LaB) source can be upgraded with a sub-200-keV DED for high-resolution cryo-EM. Using this imaging configuration, we successfully obtained a 2.65 Å reconstruction for apoferritin, 4.33 Å for 64-kDa hemoglobin, and 4.4 Å for an asymmetric 153kDa membrane protein GPCR. All results were achieved using standard automated data collection with SerialEM, demonstrating the feasibility to collect large cryo-EM datasets with a side-entry cryo-holder. These results showcase a widely accessible solution to obtaining interpretable cryo-EM structures at low cost and contribute to the "democratization" of cryo-EM.
External linksSci Adv / PubMed:39752481 / PubMed Central
MethodsEM (single particle)
Resolution2.65 Å
Structure data

EMDB-44745: 2.65 Angstrom Apoferritin single particle cryo-EM reconstruction from a standard 120-keV LaB6 electron microscope (Tecnai G2 spirit) fitted with Gatan Alpine camera (a 100keV optimised direct electron detector).
Method: EM (single particle) / Resolution: 2.65 Å

Source
  • Mus musculus (house mouse)

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