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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. Furthermore, 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.

Authors:
 [1];  [2];  [2];  [1];  [3];  [4];  [5];  [1];  [6];  [1];  [1];  [2];  [1];  [7];  [1];  [8];  [9];  [1];  [10];  [6] more »;  [6];  [11];  [11];  [12];  [12];  [13];  [13];  [14];  [4];  [15];  [16];  [16];  [17];  [4];  [4];  [4];  [4];  [12];  [18];  [6];  [3];  [6];  [11];  [19];  [1] « less
  1. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)
  2. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Univ. of Hamburg, Hamburg (Germany)
  3. Stanford Univ., Stanford, CA (United States)
  4. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  5. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Max-Planck-Institute for Biochemistry, Munich (Germany)
  6. Arizona State Univ., Tempe, AZ (United States)
  7. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Centre for Ultrafast Imaging, Hamburg (Germany)
  8. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); European XFEL GmbH, Hamburg (Germany)
  9. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Univ. of Hamburg, Hamburg (Germany); Max-Planck Institute for the Structure and Dynamics of Matter, Hamburg (Germany)
  10. Arizona State Univ., Tempe, AZ (United States); Paul Scherrer Inst. (PSI), Villigen (Switzerland)
  11. Cornell Univ., Ithaca, NY (United States)
  12. Charite - Univ. Berlin, Berlin (Germany)
  13. Technische Univ. Berlin, Berlin (Germany)
  14. Hauptman-Woodward Medical Research Institute, Buffalo, NY (United States)
  15. Univ. of Nova Gorica, Nova Gorica (Slovenia); Institute of Metals and Technology, Ljubljana (Slovenia)
  16. Institute of Metals and Technology, Ljubljana (Slovenia)
  17. Helmholtz-Zentrum Geesthacht, Geesthacht (Germany)
  18. Univ. of Minnesota, Minneapolis, MN (United States)
  19. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Univ. of Hamburg, Hamburg (Germany); Univ. of Minnesota, Minneapolis, MN (United States)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1352549
Grant/Contract Number:  
AC02-76SF00515
Resource Type:
Accepted Manuscript
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Volume: 7; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSICS; fluid dynamics; nanocrystallography

Citation Formats

Oberthuer, Dominik, Knoška, Juraj, Wiedorn, Max O., Beyerlein, Kenneth R., Bushnell, David A., Kovaleva, Elena G., Heymann, Michael, Gumprecht, Lars, Kirian, Richard A., Barty, Anton, Mariani, Valerio, Tolstikova, Aleksandra, Adriano, Luigi, Awel, Salah, Barthelmess, Miriam, Dörner, Katerina, Xavier, P. Lourdu, Yefanov, Oleksandr, James, Daniel R., Nelson, Garrett, Wang, Dingjie, Calvey, George, Chen, Yujie, Schmidt, Andrea, Szczepek, Michael, Frielingsdorf, Stefan, Lenz, Oliver, Snell, Edward, Robinson, Philip J., Šarler, Božidar, Belšak, Grega, Maček, Marjan, Wilde, Fabian, Aquila, Andrew, Boutet, Sébastien, Liang, Mengning, Hunter, Mark S., Scheerer, Patrick, Lipscomb, John D., Weierstall, Uwe, Kornberg, Roger D., Spence, John C. H., Pollack, Lois, Chapman, Henry N., and Bajt, Saša. Double-flow focused liquid injector for efficient serial femtosecond crystallography. United States: N. p., 2017. Web. doi:10.1038/srep44628.
Oberthuer, Dominik, Knoška, Juraj, Wiedorn, Max O., Beyerlein, Kenneth R., Bushnell, David A., Kovaleva, Elena G., Heymann, Michael, Gumprecht, Lars, Kirian, Richard A., Barty, Anton, Mariani, Valerio, Tolstikova, Aleksandra, Adriano, Luigi, Awel, Salah, Barthelmess, Miriam, Dörner, Katerina, Xavier, P. Lourdu, Yefanov, Oleksandr, James, Daniel R., Nelson, Garrett, Wang, Dingjie, Calvey, George, Chen, Yujie, Schmidt, Andrea, Szczepek, Michael, Frielingsdorf, Stefan, Lenz, Oliver, Snell, Edward, Robinson, Philip J., Šarler, Božidar, Belšak, Grega, Maček, Marjan, Wilde, Fabian, Aquila, Andrew, Boutet, Sébastien, Liang, Mengning, Hunter, Mark S., Scheerer, Patrick, Lipscomb, John D., Weierstall, Uwe, Kornberg, Roger D., Spence, John C. H., Pollack, Lois, Chapman, Henry N., & Bajt, Saša. Double-flow focused liquid injector for efficient serial femtosecond crystallography. United States. doi:10.1038/srep44628.
Oberthuer, Dominik, Knoška, Juraj, Wiedorn, Max O., Beyerlein, Kenneth R., Bushnell, David A., Kovaleva, Elena G., Heymann, Michael, Gumprecht, Lars, Kirian, Richard A., Barty, Anton, Mariani, Valerio, Tolstikova, Aleksandra, Adriano, Luigi, Awel, Salah, Barthelmess, Miriam, Dörner, Katerina, Xavier, P. Lourdu, Yefanov, Oleksandr, James, Daniel R., Nelson, Garrett, Wang, Dingjie, Calvey, George, Chen, Yujie, Schmidt, Andrea, Szczepek, Michael, Frielingsdorf, Stefan, Lenz, Oliver, Snell, Edward, Robinson, Philip J., Šarler, Božidar, Belšak, Grega, Maček, Marjan, Wilde, Fabian, Aquila, Andrew, Boutet, Sébastien, Liang, Mengning, Hunter, Mark S., Scheerer, Patrick, Lipscomb, John D., Weierstall, Uwe, Kornberg, Roger D., Spence, John C. H., Pollack, Lois, Chapman, Henry N., and Bajt, Saša. Thu . "Double-flow focused liquid injector for efficient serial femtosecond crystallography". United States. doi:10.1038/srep44628. https://www.osti.gov/servlets/purl/1352549.
@article{osti_1352549,
title = {Double-flow focused liquid injector for efficient serial femtosecond crystallography},
author = {Oberthuer, Dominik and Knoška, Juraj and Wiedorn, Max O. and Beyerlein, Kenneth R. and Bushnell, David A. and Kovaleva, Elena G. and Heymann, Michael and Gumprecht, Lars and Kirian, Richard A. and Barty, Anton and Mariani, Valerio and Tolstikova, Aleksandra and Adriano, Luigi and Awel, Salah and Barthelmess, Miriam and Dörner, Katerina and Xavier, P. Lourdu and Yefanov, Oleksandr and James, Daniel R. and Nelson, Garrett and Wang, Dingjie and Calvey, George and Chen, Yujie and Schmidt, Andrea and Szczepek, Michael and Frielingsdorf, Stefan and Lenz, Oliver and Snell, Edward and Robinson, Philip J. and Šarler, Božidar and Belšak, Grega and Maček, Marjan and Wilde, Fabian and Aquila, Andrew and Boutet, Sébastien and Liang, Mengning and Hunter, Mark S. and Scheerer, Patrick and Lipscomb, John D. and Weierstall, Uwe and Kornberg, Roger D. and Spence, John C. H. and Pollack, Lois and Chapman, Henry N. and Bajt, Saša},
abstractNote = {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. Furthermore, 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.},
doi = {10.1038/srep44628},
journal = {Scientific Reports},
number = ,
volume = 7,
place = {United States},
year = {2017},
month = {3}
}

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Cited by: 15 works
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Figures / Tables:

Figure 1 Figure 1: (a) Diagram of a SFX experiment at LCLS using a double flow-focusing nozzle (DFFN). Crystals of RNA polymerase II were injected in their crystallization buffer (inner green stream) into the sheath jet formed by ethanol (middle blue stream) through a 40 μm ID capillary. The sheath liquid ismore » accelerated by Helium gas (outer blank channels). A detailed view of the DFFN used in this experiment is shown in (b) and in (c) a radiograph of a working DFFN, in which the inner jet (4 M KI) can be clearly seen within the sheath jet (ethanol).« less

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    Works referencing / citing this record:

    On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies
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    Aerodynamically stabilized Taylor cone jets
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    Enzyme intermediates captured “on the fly” by mix-and-inject serial crystallography
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    Membrane protein megahertz crystallography at the European XFEL
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      Figures/Tables have been extracted from DOE-funded journal article accepted manuscripts.