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

DOI: 10.1038/s41467-020-18156-7 DOI Help

Authors: Austin Echelmeier (Arizona State University) , Jorvani Cruz Villarreal (Arizona State University) , Marc Messerschmidt (Arizona State University; European XFEL) , 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; Universität Hamburg) , J. Domingo Meza-Aguilar (European XFEL) , Dominik Oberthuer (Deutsches Elektronen-Synchrotron DESY) , Max O. Wiedorn (Universität Hamburg; Deutsches Elektronen-Synchrotron) , Holger Fleckenstein (Deutsches Elektronen-Synchrotron) , Derek Mendez (Arizona State University) , Juraj Knoška (Deutsches Elektronen-Synchrotron; Universität Hamburg) , Jose M. Martin-Garcia (Arizona State University) , Hao Hu (Arizona State University) , Stella Lisova (Arizona State University) , Aschkai Allahgoli (Deutsches Elektronen-Synchrotron) , Yaroslav Gevorkov (Deutsches Elektronen-Synchrotron; Hamburg University of Technology) , Kartik Ayyer (Deutsches Elektronen-Synchrotron) , Steve Aplin (Deutsches Elektronen-Synchrotron) , Helen M. Ginn (University of Oxford; Diamond Light Source) , 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 DESY) , Jennifer Poehlsen (Deutsches Elektronen-Synchrotron) , Ulrich Trunk (Deutsches Elektronen-Synchrotron) , Davide Mezza (Paul Scherrer Institute) , Bernd Schmitt (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; Deutsches Elektronen-Synchrotron) , Klaus Giewekemeyer (European XFEL) , Adam Round (European XFEL; Keele University) , 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; La Trobe University) , Uwe Weierstall (Arizona State University) , John C. H. Spence (Arizona State University) , Henry N. Chapman (Universität Hamburg; Deutsches Elektronen-Synchrotron) , Nadia Zatsepin (Arizona State University; La Trobe University) , Petra Fromme (Arizona State University) , Richard A. Kirian (Arizona State University) , Alexandra Ros (Arizona State University)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Communications , VOL 11

State: Published (Approved)
Published: September 2020

Open Access Open Access

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.

Journal Keywords: Microfluidics; Nanocrystallography; Structural biology; X-ray crystallography

Subject Areas: Technique Development, Biology and Bio-materials

Facility: European XFEL

Added On: 20/09/2020 16:10


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Technique Development - Life Sciences & Biotech Structural biology Life Sciences & Biotech

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