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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].

SUBMITTER: Oberthuer D 

PROVIDER: S-EPMC5353652 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature

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

Oberthuer Dominik D   Knoška Juraj J   Wiedorn Max O MO   Beyerlein Kenneth R KR   Bushnell David A DA   Kovaleva Elena G EG   Heymann Michael M   Gumprecht Lars L   Kirian Richard A RA   Barty Anton A   Mariani Valerio V   Tolstikova Aleksandra A   Adriano Luigi L   Awel Salah S   Barthelmess Miriam M   Dörner Katerina K   Xavier P Lourdu PL   Yefanov Oleksandr O   James Daniel R DR   Nelson Garrett G   Wang Dingjie D   Calvey George G   Chen Yujie Y   Schmidt Andrea A   Szczepek Michael M   Frielingsdorf Stefan S   Lenz Oliver O   Snell Edward E   Robinson Philip J PJ   Šarler Božidar B   Belšak Grega G   Maček Marjan M   Wilde Fabian F   Aquila Andrew A   Boutet Sébastien S   Liang Mengning M   Hunter Mark S MS   Scheerer Patrick P   Lipscomb John D JD   Weierstall Uwe U   Kornberg Roger D RD   Spence John C H JC   Pollack Lois L   Chapman Henry N HN   Bajt Saša S  

Scientific reports 20170316

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 det  ...[more]

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