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.
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- Double-flow Focused Liquid Injector for Efficient Serial Femtosecond Crystallography
- Oberthuer, Dominik (Author)
- Knoska, Juraj (Author)
- Wiedorn, Max O. (Author)
- Beyerlein, Kenneth R. (Author)
- Bushnell, David A. (Author)
- Kovaleva, Elena G. (Author)
- Heymann, Michael (Author)
- Gumprecht, Lars (Author)
- Kirian, Richard (Author)
- Barty, Anton (Author)
- Mariani, Valerio (Author)
- Tolstikova, Aleksandra (Author)
- Adriano, Luigi (Author)
- Awel, Salah (Author)
- Barthelmess, Miriam (Author)
- Dorner, Katerina (Author)
- Xavier, P. Lourdu (Author)
- Yefanov, Oleksandr (Author)
- James, Daniel (Author)
- Nelson, Garrett (Author)
- Wang, Dingjie (Author)
- Calvey, George (Author)
- Chen, Yujie (Author)
- Schmidt, Andrea (Author)
- Szczepek, Michael (Author)
- Frielingsdorf, Stefan (Author)
- Lenz, Oliver (Author)
- Snell, Edward (Author)
- Robinson, Philip J. (Author)
- Sarler, Bozidar (Author)
- Belsak, Grega (Author)
- Macek, Marjan (Author)
- Wilde, Fabian (Author)
- Aquila, Andrew (Author)
- Boutet, Sebastien (Author)
- Liang, Mengning (Author)
- Hunter, Mark S. (Author)
- Scheerer, Patrick (Author)
- Lipscomb, John D. (Author)
- Weierstall, Uwe (Author)
- Kornberg, Roger D. (Author)
- Spence, John (Author)
- Pollack, Lois (Author)
- Chapman, Henry N. (Author)
- Bajt, Sasa (Author)
- College of Liberal Arts and Sciences (Contributor)
- Department of Physics (Contributor)
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Digital object identifier: 10.1038/srep44628
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Identifier TypeInternational standard serial numberIdentifier Value2045-2322
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The final version of this article, as published in Scientific Reports, can be viewed online at: https://www.nature.com/articles/srep44628
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Oberthuer, D., Knoška, J., Wiedorn, M. O., Beyerlein, K. R., Bushnell, D. A., Kovaleva, E. G., . . . Bajt, S. (2017). Double-flow focused liquid injector for efficient serial femtosecond crystallography. Scientific Reports, 7, 44628. doi:10.1038/srep44628