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PDBsum entry 4z98
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References listed in PDB file
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Key reference
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Title
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A high-Transparency, Micro-Patternable chip for X-Ray diffraction analysis of microcrystals under native growth conditions.
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Authors
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T.D.Murray,
A.Y.Lyubimov,
C.M.Ogata,
H.Vo,
M.Uervirojnangkoorn,
A.T.Brunger,
J.M.Berger.
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Ref.
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Acta Crystallogr D Biol Crystallogr, 2015,
71,
1987-1997.
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PubMed id
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Abstract
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Microcrystals present a significant impediment to the determination of
macromolecular structures by X-ray diffraction methods. Although microfocus
synchrotron beamlines and X-ray free-electron lasers (XFELs) can enable the
collection of interpretable diffraction data from microcrystals, there is a need
for efficient methods of harvesting small volumes (<2 µl) of microcrystals
grown under common laboratory formats and delivering them to an X-ray beam
source under native growth conditions. One approach that shows promise in
overcoming the challenges intrinsic to microcrystal analysis is to pair
so-called `fixed-target' sample-delivery devices with microbeam-based X-ray
diffraction methods. However, to record weak diffraction patterns it is
necessary to fabricate devices from X-ray-transparent materials that minimize
background scattering. Presented here is the design of a new micro-diffraction
device consisting of three layers fabricated from silicon nitride, photoresist
and polyimide film. The chip features low X-ray scattering and X-ray absorption
properties, and uses a customizable blend of hydrophobic and hydrophilic surface
patterns to help localize microcrystals to defined regions. Microcrystals in
their native growth conditions can be loaded into the chips with a standard
pipette, allowing data collection at room temperature. Diffraction data
collected from hen egg-white lysozyme microcrystals (10-15 µm) loaded into
the chips yielded a complete, high-resolution (<1.6 Å) data set sufficient
to determine a high-quality structure by molecular replacement. The features of
the chip allow the rapid and user-friendly analysis of microcrystals grown under
virtually any laboratory format at microfocus synchrotron beamlines and XFELs.
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