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

By Austin Echelmeier, Jorvani Cruz Villarreal, Marc Messerschmidt1, Daihyun Kim, Jesse David Coe1, Darren Gary Thifault1, Sabine Botha1, Ana Egatz-Gomez2, Sahir Ilyas Gandhi1, Gerrit Brehm, Chelsie E Conrad1, Debra T. Hansen, Caleb Alexander Madsen3, Saša Bajt, J. Domingo Meza-Aguilar, Dominik Oberthür, Max Oliver Wiedorn4, Holger Fleckenstein, Derek Mendez1, 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 Weierstall1, John Spence1, Henry Chapman4, Nadia Zatsepin1, Petra Fromme1, Richard Kirian4, Alexandra Ros1

1. Arizona State University 2. Center for Applied Structural Discovery 3. BioXFEL Undergraduate Summer Intern 4. Center for Free-Electron Laser Science

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Category

BioXFEL Publications

Published on

Type

journal-article

Author

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

Citation

Echelmeier, A. et al., 2020. Segmented flow generator for serial crystallography at the European X-ray free electron laser. Nature Communications, 11(1). Available at: http://dx.doi.org/10.1038/s41467-020-18156-7.

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.

DOI

http://dx.doi.org/10.1038/s41467-020-18156-7

Funding

NSF-STC Biology with X-ray Lasers (NSF-1231306)