National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R01GM098621
United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R01GM116942
United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R35GM136258
United States
Citation
Journal: J Struct Biol X / Year: 2021 Title: Experimental evaluation of super-resolution imaging and magnification choice in single-particle cryo-EM. Authors: J Ryan Feathers / Katherine A Spoth / J Christopher Fromme / Abstract: The resolution of cryo-EM reconstructions is fundamentally limited by the Nyquist frequency, which is half the sampling frequency of the detector and depends upon the magnification used. In ...The resolution of cryo-EM reconstructions is fundamentally limited by the Nyquist frequency, which is half the sampling frequency of the detector and depends upon the magnification used. In principle, super-resolution imaging should enable reconstructions to surpass the physical Nyquist limit by increasing sampling frequency, yet there are few reports of reconstructions that do so. Here we directly examine the contribution of super-resolution information, obtained with the K3 direct electron detector using a 2-condenser microscope, to single-particle cryo-EM reconstructions surpassing the physical Nyquist limit. We also present a comparative analysis of a sample imaged at four different magnifications. This analysis demonstrates that lower magnifications can be beneficial, despite the loss of higher resolution signal, due to the increased number of particle images obtained. To highlight the potential utility of lower magnification data collection, we produced a 3.5 Å reconstruction of jack bean urease with particles from a single micrograph.
EMPIAR-10547 (Title: Jack bean urease imaged at 49kX nominal magnification Data size: 169.3 / Data #1: Super resolution movies [micrographs - multiframe])
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