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).
Map data
B-Factor Sharpened Map
Sample
Complex: Apoferritin
Keywords
Apoferritin / High resolution / LaB6 / Tecnai. / METAL TRANSPORT
Biological species
Mus musculus (house mouse)
Method
single particle reconstruction / cryo EM / Resolution: 2.65 Å
National Health and Medical Research Council (NHMRC, Australia)
2010
Australia
Chan Zuckerberg Initiative
DAF2021-225399
United States
Citation
Journal: Sci Adv / Year: 2025 Title: High-resolution cryo-EM using a common LaB 120-keV electron microscope equipped with a sub-200-keV direct electron detector. Authors: Hariprasad Venugopal / Jesse Mobbs / Cyntia Taveneau / Daniel R Fox / Ziva Vuckovic / Sahil Gulati / Gavin Knott / Rhys Grinter / David Thal / Stephen Mick / Cory Czarnik / Georg Ramm / Abstract: 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- ...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.
pH: 8 / Details: 50 mM Tris-HCl (pH 8.0), 150 mM NaCl
Vitrification
Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV
-
Electron microscopy
Microscope
FEI TECNAI SPIRIT
Image recording
Film or detector model: GATAN ALPINE (2.3k x 3.2k) / Digitization - Dimensions - Width: 2304 pixel / Digitization - Dimensions - Height: 3240 pixel / Number grids imaged: 1 / Number real images: 923 / Average exposure time: 6.55 sec. / Average electron dose: 50.3 e/Å2
Electron beam
Acceleration voltage: 120 kV / Electron source: LAB6
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