Cryogenic x-ray diffraction microscopy utilizing high-pressure cryopreservation

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 environment...

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Published inPhysical review. E, Statistical, nonlinear, and soft matter physics Vol. 90; no. 4; p. 042713
Main Authors Lima, Enju, Chushkin, Yuriy, van der Linden, Peter, Kim, Chae Un, Zontone, Federico, Carpentier, Philippe, Gruner, Sol M, Pernot, Petra
Format Journal Article
LanguageEnglish
Published United States 15.10.2014
Subjects
Cryopreservation - methods
Deinococcus
Ice
Image Processing, Computer-Assisted
Microscopy - methods
Pressure
Scattering, Small Angle
X-Ray Diffraction - methods
X-Rays
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Summary: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.
ISSN:1550-2376
DOI:10.1103/PhysRevE.90.042713