Cryo-EM with sub–1 Å specimen movement

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Science  09 Oct 2020:
Vol. 370, Issue 6513, pp. 223-226
DOI: 10.1126/science.abb7927

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Going for the gold

Single-particle cryogenic electron microscopy (cryo-EM) has become a go-to technique for structural biologists. Although data-processing and reconstruction methods have improved, innovations in sample preparation and data collection are essential to reliably achieve high-resolution reconstructions while also reducing the amount of time required per structure. Naydenova et al. tackled the issue of electron beam–induced particle movement, a major source of information loss, by designing a gold sample support that prevents buckling of the extremely thin layer of ice in which the particles are suspended (see Perspective by Rapp and Carragher). The negligible particle displacement permits extrapolation to “zero exposure” structure factors, revealing features typically lost in cryo-EM structures. Far fewer particles per unit resolution are required, which greatly accelerates structure determination, especially at high resolution.

Science, this issue p. 223; see also p. 171


Most information loss in cryogenic electron microscopy (cryo-EM) stems from particle movement during imaging, which remains poorly understood. We show that this movement is caused by buckling and subsequent deformation of the suspended ice, with a threshold that depends directly on the shape of the frozen water layer set by the support foil. We describe a specimen support design that eliminates buckling and reduces electron beam–induced particle movement to less than 1 angstrom. The design allows precise foil tracking during imaging with high-speed detectors, thereby lessening demands on cryostage precision and stability. It includes a maximal density of holes, which increases throughput in automated cryo-EM without degrading data quality. Movement-free imaging allows extrapolation to a three-dimensional map of the specimen at zero electron exposure, before the onset of radiation damage.

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