Segmented flow generator for serial crystallography at the European X-ray free electron laser

Austin Echelmeier, Jorvani Cruz Villarreal, Marc Messerschmidt, Daihyun Kim, Jesse D. Coe, Darren Thifault, Sabine Botha, Ana Egatz-Gomez, Sahir Gandhi, Gerrit Brehm, Chelsie E. Conrad, Debra T. Hansen, Caleb Madsen, Saša Bajt, J. Domingo Meza-Aguilar, Dominik Oberthür, Max O. Wiedorn, Holger Fleckenstein, Derek Mendez, Juraj Knoška, Jose M. Martin-Garcia, Hao Hu, Stella Lisova, Aschkan Allahgholi, Yaroslav Gevorkov, Kartik Ayyer, Steve Aplin, Helen Mary Ginn, Heinz Graafsma, Andrew J. Morgan, Dominic Greiffenberg, Alexander Klujev, Torsten Laurus, Jennifer Poehlsen, Ulrich Trunk, Davide Mezza, Bernd Schmidt, Manuela Kuhn, Raimund Fromme, Jolanta Sztuk-Dambietz, Natascha Raab, Steffen Hauf, Alessandro Silenzi, Thomas Michelat, Chen Xu, Cyril Danilevski, Andrea Parenti, Leonce Mekinda, Britta Weinhausen, Grant Mills, Patrik Vagovic, Yoonhee Kim, Henry Kirkwood, Richard Bean, Johan Bielecki, Stephan Stern, Klaus Giewekemeyer, Adam R. Round, Joachim Schulz, Katerina Dörner, Thomas D. Grant, Valerio Mariani, Anton Barty, Adrian P. Mancuso, Uwe Weierstall, John C. H. Spence, Henry N. Chapman, Nadia Zatsepin, Petra Fromme, Richard A. Kirian and Alexandra Ros ()
Additional contact information
Austin Echelmeier: Arizona State University
Jorvani Cruz Villarreal: Arizona State University
Marc Messerschmidt: Arizona State University
Daihyun Kim: Arizona State University
Jesse D. Coe: Arizona State University
Darren Thifault: Arizona State University
Sabine Botha: Arizona State University
Ana Egatz-Gomez: Arizona State University
Sahir Gandhi: Arizona State University
Gerrit Brehm: University of Göttingen
Chelsie E. Conrad: Arizona State University
Debra T. Hansen: Arizona State University
Caleb Madsen: Arizona State University
Saša Bajt: Deutsches Elektronen-Synchrotron
J. Domingo Meza-Aguilar: European XFEL
Dominik Oberthür: Deutsches Elektronen-Synchrotron
Max O. Wiedorn: Universität Hamburg
Holger Fleckenstein: Deutsches Elektronen-Synchrotron
Derek Mendez: Arizona State University
Juraj Knoška: Deutsches Elektronen-Synchrotron
Jose M. Martin-Garcia: Arizona State University
Hao Hu: Arizona State University
Stella Lisova: Arizona State University
Aschkan Allahgholi: Deutsches Elektronen-Synchrotron
Yaroslav Gevorkov: Deutsches Elektronen-Synchrotron
Kartik Ayyer: Deutsches Elektronen-Synchrotron
Steve Aplin: Deutsches Elektronen-Synchrotron
Helen Mary Ginn: University of Oxford
Heinz Graafsma: Deutsches Elektronen-Synchrotron
Andrew J. Morgan: Deutsches Elektronen-Synchrotron
Dominic Greiffenberg: Paul Scherrer Institute
Alexander Klujev: Deutsches Elektronen-Synchrotron
Torsten Laurus: Deutsches Elektronen-Synchrotron
Jennifer Poehlsen: Deutsches Elektronen-Synchrotron
Ulrich Trunk: Deutsches Elektronen-Synchrotron
Davide Mezza: Paul Scherrer Institute
Bernd Schmidt: Paul Scherrer Institute
Manuela Kuhn: Deutsches Elektronen-Synchrotron
Raimund Fromme: Arizona State University
Jolanta Sztuk-Dambietz: European XFEL
Natascha Raab: European XFEL
Steffen Hauf: European XFEL
Alessandro Silenzi: European XFEL
Thomas Michelat: European XFEL
Chen Xu: European XFEL
Cyril Danilevski: European XFEL
Andrea Parenti: European XFEL
Leonce Mekinda: European XFEL
Britta Weinhausen: European XFEL
Grant Mills: European XFEL
Patrik Vagovic: European XFEL
Yoonhee Kim: European XFEL
Henry Kirkwood: European XFEL
Richard Bean: European XFEL
Johan Bielecki: European XFEL
Stephan Stern: European XFEL
Klaus Giewekemeyer: European XFEL
Adam R. Round: European XFEL
Joachim Schulz: European XFEL
Katerina Dörner: European XFEL
Thomas D. Grant: SUNY University at Buffalo
Valerio Mariani: Deutsches Elektronen-Synchrotron
Anton Barty: Deutsches Elektronen-Synchrotron
Adrian P. Mancuso: European XFEL
Uwe Weierstall: Arizona State University
John C. H. Spence: Arizona State University
Henry N. Chapman: Universität Hamburg
Nadia Zatsepin: Arizona State University
Petra Fromme: Arizona State University
Richard A. Kirian: Arizona State University
Alexandra Ros: Arizona State University

Nature Communications, 2020, vol. 11, issue 1, 1-10

Abstract: Abstract Serial femtosecond crystallography (SFX) with X-ray free electron lasers (XFELs) allows structure determination of membrane proteins and time-resolved crystallography. Common liquid sample delivery continuously jets the protein crystal suspension into the path of the XFEL, wasting a vast amount of sample due to the pulsed nature of all current XFEL sources. The European XFEL (EuXFEL) delivers femtosecond (fs) X-ray pulses in trains spaced 100 ms apart whereas pulses within trains are currently separated by 889 ns. Therefore, continuous sample delivery via fast jets wastes >99% of sample. Here, we introduce a microfluidic device delivering crystal laden droplets segmented with an immiscible oil reducing sample waste and demonstrate droplet injection at the EuXFEL compatible with high pressure liquid delivery of an SFX experiment. While achieving ~60% reduction in sample waste, we determine the structure of the enzyme 3-deoxy-D-manno-octulosonate-8-phosphate synthase from microcrystals delivered in droplets revealing distinct structural features not previously reported.

Date: 2020
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Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-18156-7

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DOI: 10.1038/s41467-020-18156-7

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