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  • Title: Cryogenic x-ray diffraction microscopy utilizing high-pressure cryopreservation.
    Author: Lima E, Chushkin Y, van der Linden P, Kim CU, Zontone F, Carpentier P, Gruner SM, Pernot P.
    Journal: Phys Rev E Stat Nonlin Soft Matter Phys; 2014 Oct; 90(4):042713. PubMed ID: 25375529.
    Abstract:
    We present cryo x-ray diffraction microscopy of high-pressure-cryofixed bacteria and report high-convergence imaging with multiple image reconstructions. Hydrated D. radiodurans cells were cryofixed at 200 MPa pressure into ∼10-μm-thick water layers and their unstained, hydrated cellular environments were imaged by phasing diffraction patterns, reaching sub-30-nm resolutions with hard x-rays. Comparisons were made with conventional ambient-pressure-cryofixed samples, with respect to both coherent small-angle x-ray scattering and the image reconstruction. The results show a correlation between the level of background ice signal and phasing convergence, suggesting that phasing difficulties with frozen-hydrated specimens may be caused by high-background ice scattering.
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