title: Double-flow focused liquid injector for efficient serial femtosecond crystallography.
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].
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.
authors.forename: Dominik
authors.initials: D
authors.lastname: Oberthuer
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.|Department of Physics, University of Hamburg, Luruper Chaussee 149, 22761 Hamburg, Germany.
authors.forename: Juraj
authors.initials: J
authors.lastname: Knoška
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.|Department of Physics, University of Hamburg, Luruper Chaussee 149, 22761 Hamburg, Germany.
authors.forename: Max O
authors.initials: MO
authors.lastname: Wiedorn
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.
authors.forename: Kenneth R
authors.initials: KR
authors.lastname: Beyerlein
authors.affiliation: Department of Structural Biology, Stanford University, School of Medicine, Stanford, California 94305, USA.
authors.forename: David A
authors.initials: DA
authors.lastname: Bushnell
authors.affiliation: SSRL, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Elena G
authors.initials: EG
authors.lastname: Kovaleva
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Heymann
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.
authors.forename: Lars
authors.initials: L
authors.lastname: Gumprecht
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ, USA.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Kirian
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.
authors.forename: Anton
authors.initials: A
authors.lastname: Barty
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.
authors.forename: Valerio
authors.initials: V
authors.lastname: Mariani
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.|Department of Physics, University of Hamburg, Luruper Chaussee 149, 22761 Hamburg, Germany.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Tolstikova
authors.affiliation: Photon Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.
authors.forename: Luigi
authors.initials: L
authors.lastname: Adriano
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.|Centre for Ultrafast Imaging, Luruper Chaussee 149, 22761 Hamburg, Germany.
authors.forename: Salah
authors.initials: S
authors.lastname: Awel
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.
authors.forename: Miriam
authors.initials: M
authors.lastname: Barthelmess
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.
authors.forename: Katerina
authors.initials: K
authors.lastname: Dörner
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.|Department of Physics, University of Hamburg, Luruper Chaussee 149, 22761 Hamburg, Germany.|IMPRS-UFAST, Max-Planck Institute for the Structure and Dynamics of Matter, 22675 Hamburg, Germany.
authors.forename: P Lourdu
authors.initials: PL
authors.lastname: Xavier
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.
authors.forename: Oleksandr
authors.initials: O
authors.lastname: Yefanov
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ, USA.
authors.forename: Daniel R
authors.initials: DR
authors.lastname: James
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ, USA.
authors.forename: Garrett
authors.initials: G
authors.lastname: Nelson
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ, USA.
authors.forename: Dingjie
authors.initials: D
authors.lastname: Wang
authors.affiliation: School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853, USA.
authors.forename: George
authors.initials: G
authors.lastname: Calvey
authors.affiliation: School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853, USA.
authors.forename: Yujie
authors.initials: Y
authors.lastname: Chen
authors.forename: Andrea
authors.initials: A
authors.lastname: Schmidt
authors.forename: Michael
authors.initials: M
authors.lastname: Szczepek
authors.affiliation: Institut für Chemie, Sekr. PC14, Technische Universität Berlin, Straße des 17, Juni 135, 10623 Berlin, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Frielingsdorf
authors.affiliation: Institut für Chemie, Sekr. PC14, Technische Universität Berlin, Straße des 17, Juni 135, 10623 Berlin, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Lenz
authors.affiliation: Hauptman-Woodward Medical Research Institute, 700 Ellicott Street, Buffalo, New York 14203, USA.
authors.forename: Edward
authors.initials: E
authors.lastname: Snell
authors.affiliation: SSRL, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Robinson
authors.affiliation: Laboratory for Multiphase Processes, University of Nova Gorica, Vipavska 13, SI-5000 Nova Gorica, Slovenia.|Laboratory for Simulation of Materials and Processes, Institute of Metals and Technology, Lepi pot 11, SI-1000 Ljubljana, Slovenia.
authors.forename: Božidar
authors.initials: B
authors.lastname: Šarler
authors.affiliation: Laboratory for Simulation of Materials and Processes, Institute of Metals and Technology, Lepi pot 11, SI-1000 Ljubljana, Slovenia.
authors.forename: Grega
authors.initials: G
authors.lastname: Belšak
authors.affiliation: Laboratory for Simulation of Materials and Processes, Institute of Metals and Technology, Lepi pot 11, SI-1000 Ljubljana, Slovenia.
authors.forename: Marjan
authors.initials: M
authors.lastname: Maček
authors.affiliation: Helmholtz-Zentrum Geesthacht, Max-Planck-Straße 1, 21502 Geesthacht, Germany.
authors.forename: Fabian
authors.initials: F
authors.lastname: Wilde
authors.affiliation: LCLS, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Aquila
authors.affiliation: LCLS, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Boutet
authors.affiliation: LCLS, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Mengning
authors.initials: M
authors.lastname: Liang
authors.affiliation: LCLS, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Mark S
authors.initials: MS
authors.lastname: Hunter
authors.forename: Patrick
authors.initials: P
authors.lastname: Scheerer
authors.affiliation: Department of Biochemistry, Molecular Biology &Biophysics, University of Minnesota, Minneapolis, Minnesota 55455, USA.
authors.forename: John D
authors.initials: JD
authors.lastname: Lipscomb
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ, USA.
authors.forename: Uwe
authors.initials: U
authors.lastname: Weierstall
authors.affiliation: Department of Structural Biology, Stanford University, School of Medicine, Stanford, California 94305, USA.
authors.forename: Roger D
authors.initials: RD
authors.lastname: Kornberg
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ, USA.
authors.forename: John C H
authors.initials: JC
authors.lastname: Spence
authors.affiliation: School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853, USA.
authors.forename: Lois
authors.initials: L
authors.lastname: Pollack
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.|Department of Physics, University of Hamburg, Luruper Chaussee 149, 22761 Hamburg, Germany.|Centre for Ultrafast Imaging, Luruper Chaussee 149, 22761 Hamburg, Germany.
authors.forename: Henry N
authors.initials: HN
authors.lastname: Chapman
authors.affiliation: Photon Science, Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg, Germany.
authors.forename: Saša
authors.initials: S
authors.lastname: Bajt
mesh_terms: D003198:Computer Simulation; D003461:Crystallography / Q000295:instrumentation*; D012319:RNA Polymerase II / Q000737:chemistry; D012212:Rheology / Q000295:instrumentation*; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013696:Temperature; D013997:Time Factors; D014961:X-Ray Diffraction
chemical_list: D012319:RNA Polymerase II