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Sci Rep. 2017 Mar 16;7:44628. doi: 10.1038/srep44628.

Double-flow focused liquid injector for efficient serial femtosecond crystallography.

Author information

1
Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.
2
Department of Physics, University of Hamburg, Luruper Chaussee 149, 22761 Hamburg, Germany.
3
Department of Structural Biology, Stanford University, School of Medicine, Stanford, California 94305, USA.
4
SSRL, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
5
Department of Physics, Arizona State University, Tempe, AZ, USA.
6
Photon Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.
7
Centre for Ultrafast Imaging, Luruper Chaussee 149, 22761 Hamburg, Germany.
8
IMPRS-UFAST, Max-Planck Institute for the Structure and Dynamics of Matter, 22675 Hamburg, Germany.
9
School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853, USA.
10
Institut für Chemie, Sekr. PC14, Technische Universität Berlin, Straße des 17, Juni 135, 10623 Berlin, Germany.
11
Hauptman-Woodward Medical Research Institute, 700 Ellicott Street, Buffalo, New York 14203, USA.
12
Laboratory for Multiphase Processes, University of Nova Gorica, Vipavska 13, SI-5000 Nova Gorica, Slovenia.
13
Laboratory for Simulation of Materials and Processes, Institute of Metals and Technology, Lepi pot 11, SI-1000 Ljubljana, Slovenia.
14
Helmholtz-Zentrum Geesthacht, Max-Planck-Straße 1, 21502 Geesthacht, Germany.
15
LCLS, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
16
Department of Biochemistry, Molecular Biology &Biophysics, University of Minnesota, Minneapolis, Minnesota 55455, USA.

Abstract

Serial femtosecond crystallography requires reliable and efficient delivery of fresh crystals across the beam of an X-ray free-electron laser over the course of an experiment. We introduce a double-flow focusing nozzle to meet this challenge, with significantly reduced sample consumption, while improving jet stability over previous generations of nozzles. We demonstrate its use to determine the first room-temperature structure of RNA polymerase II at high resolution, revealing new structural details. Moreover, the double-flow focusing nozzles were successfully tested with three other protein samples and the first room temperature structure of an extradiol ring-cleaving dioxygenase was solved by utilizing the improved operation and characteristics of these devices [corrected].

PMID:
28300169
PMCID:
PMC5353652
DOI:
10.1038/srep44628
[Indexed for MEDLINE]
Free PMC Article

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