Double-flow focused liquid injector for efficient serial femtosecond crystallography

Dominik Oberthuer, Juraj Knoška, Max O. Wiedorn, Kenneth R. Beyerlein, David A. Bushnell, Elena G. Kovaleva, Michael Heymann, Lars Gumprecht, Richard A. Kirian, Anton Barty, Valerio Mariani, Aleksandra Tolstikova, Luigi Adriano, Salah Awel, Miriam Barthelmess, Katerina Dörner, P. Lourdu Xavier, Oleksandr Yefanov, Daniel R. James, Garrett NelsonDingjie Wang, George Calvey, Yujie Chen, Andrea Schmidt, Michael Szczepek, Stefan Frielingsdorf, Oliver Lenz, Edward Snell, Philip J. Robinson, Böidar Šarler, Grega Belšak, Marjan Maček, Fabian Wilde, Andrew Aquila, Sébastien Boutet, Mengning Liang, Mark S. Hunter, Patrick Scheerer, John D. Lipscomb, Uwe Weierstall, Roger D. Kornberg, John C.H. Spence, Lois Pollack, Henry N. Chapman, Saša Bajt

Research output: Contribution to journalArticle

50 Scopus citations

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.

Original languageEnglish (US)
Article number44628
JournalScientific reports
Volume7
DOIs
StatePublished - Mar 16 2017

Fingerprint Dive into the research topics of 'Double-flow focused liquid injector for efficient serial femtosecond crystallography'. Together they form a unique fingerprint.

  • Cite this

    Oberthuer, D., Knoška, J., Wiedorn, M. O., Beyerlein, K. R., Bushnell, D. A., Kovaleva, E. G., Heymann, M., Gumprecht, L., Kirian, R. A., Barty, A., Mariani, V., Tolstikova, A., Adriano, L., Awel, S., Barthelmess, M., Dörner, K., Xavier, P. L., Yefanov, O., James, D. R., ... Bajt, S. (2017). Double-flow focused liquid injector for efficient serial femtosecond crystallography. Scientific reports, 7, [44628]. https://doi.org/10.1038/srep44628