1. Segmented flow generator for serial crystallography at the European X-ray free electron laser
- Author
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Steve Aplin, Nadia A. Zatsepin, Darren Thifault, Anton Barty, Jennifer Poehlsen, Manuela Kuhn, Leonce Mekinda, Jolanta Sztuk-Dambietz, Max O. Wiedorn, Andrea Parenti, Y. Gevorkov, Yoonhee Kim, Debra T. Hansen, Grant Mills, Saša Bajt, Henry Kirkwood, Davide Mezza, Katerina Dörner, Joachim Schulz, Helen M. Ginn, Alexander Klujev, Hao Hu, Valerio Mariani, Patrik Vagovic, Jesse Coe, Kartik Ayyer, Juraj Knoska, U. Trunk, Thomas D. Grant, John C. H. Spence, Austin Echelmeier, Chelsie E. Conrad, Marc Messerschmidt, Chen Xu, Thomas Michelat, Adam Round, Henry N. Chapman, Natascha Raab, Uwe Weierstall, Ana Egatz-Gomez, Klaus Giewekemeyer, Holger Fleckenstein, Dominik Oberthür, Stella Lisova, Bernd Schmidt, Raimund Fromme, Richard Bean, Petra Fromme, Torsten Laurus, Cyril Danilevski, Stephan Stern, J. Domingo Meza-Aguilar, Steffen Hauf, Adrian P. Mancuso, Alessandro Silenzi, Jorvani Cruz Villarreal, Dominic Greiffenberg, A. Allahgholi, Sahir Gandhi, Andrew J. Morgan, Gerrit Brehm, Johan Bielecki, Britta Weinhausen, Jose M. Martin-Garcia, Sabine Botha, Heinz Graafsma, Derek Mendez, Daihyun Kim, Caleb Madsen, Richard A. Kirian, and Alexandra Ros
- Subjects
0301 basic medicine ,Free electron model ,60199 Biochemistry and Cell Biology not elsewhere classified ,Materials science ,Science ,Microfluidics ,General Physics and Astronomy ,Electrons ,02 engineering and technology ,General Biochemistry, Genetics and Molecular Biology ,Article ,law.invention ,Crystal ,03 medical and health sciences ,law ,Lab-On-A-Chip Devices ,lcsh:Science ,Technik [600] ,Aldehyde-Lyases ,X-ray crystallography ,Multidisciplinary ,Crystallography ,Nanocrystallography ,Escherichia coli Proteins ,Lasers ,600: Technik ,Free-electron laser ,General Chemistry ,021001 nanoscience & nanotechnology ,Laser ,030104 developmental biology ,FOS: Biological sciences ,Femtosecond ,Hydrodynamics ,lcsh:Q ,ddc:500 ,0210 nano-technology ,Protein crystallization ,Structural biology ,ddc:600 - Abstract
Nature Communications 11(1), 4511 (2020). doi:10.1038/s41467-020-18156-7, 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., Published by Nature Publishing Group UK, [London]
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- 2023
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