ArticleNature communications2020
Segmented flow generator for serial crystallography at the European X-ray free electron laser.
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Article in Nature communications, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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16 citing papers in PubMed.
- Drop-on-fixed-target reaction initiation approach for serial and time-resolved crystallography.IUCrJ · 2026Article
- Minimized sample consumption for time-resolved serial crystallography applied to the redox cycle of human NQO1.Communications chemistry · 2026Article
- Sample delivery methods for protein X-ray crystallography with a special focus on sample consumption.Nature communications · 2025Review
- Review
- Cyclic Olefin Copolymer-Based Fixed-Target Sample Delivery Device for Protein X-ray Crystallography.Analytical chemistry · 2024Article
- Microsecond time-resolved X-ray scattering by utilizing MHz repetition rate at second-generation XFELs.Nature methods · 2024Article
- Kilohertz droplet-on-demand serial femtosecond crystallography at the European XFEL station FXE.Structural dynamics (Melville, N.Y.) · 2024Article
- Modular droplet injector for sample conservation providing new structural insight for the conformational heterogeneity in the disease-associated NQO1 enzyme.Lab on a chip · 2023Article
- Emerging Time-Resolved X-Ray Diffraction Approaches for Protein Dynamics.Annual review of biophysics · 2023Review
- Electrically stimulated droplet injector for reduced sample consumption in serial crystallography.Biophysical reports · 2022Article
- Expected resolution limits of x-ray free-electron laser single-particle imaging for realistic source and detector properties.Structural dynamics (Melville, N.Y.) · 2022Article
- Megahertz pulse trains enable multi-hit serial femtosecond crystallography experiments at X-ray free electron lasers.Nature communications · 2022Article
- Serial femtosecond crystallography.Nature reviews. Methods primers · 2022Article
- A multi-million image Serial Femtosecond Crystallography dataset collected at the European XFEL.Scientific data · 2022Article
- Beef tallow injection matrix for serial crystallography.Scientific reports · 2022Article
- Wall-free droplet microfluidics for probing biological processes by high-brilliance X-ray scattering techniques.Frontiers in molecular biosciences · 2022Review
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Authors and funding
71 authors.
Austin Echelmeier *School of Molecular Sciences, Arizona State University, Tempe, AZ, 85287-1604, USA.ORCID http://orcid.org/0000-0001-6127-3879
Jorvani Cruz Villarreal *School of Molecular Sciences, Arizona State University, Tempe, AZ, 85287-1604, USA.ORCID http://orcid.org/0000-0002-6762-7348
Marc MesserschmidtSchool of Molecular Sciences, Arizona State University, Tempe, AZ, 85287-1604, USA.
Daihyun KimSchool of Molecular Sciences, Arizona State University, Tempe, AZ, 85287-1604, USA.
Jesse D CoeSchool of Molecular Sciences, Arizona State University, Tempe, AZ, 85287-1604, USA.ORCID http://orcid.org/0000-0002-5544-320X
Darren ThifaultSchool of Molecular Sciences, Arizona State University, Tempe, AZ, 85287-1604, USA.ORCID http://orcid.org/0000-0001-9710-1307
Sabine BothaCenter for Applied Structural Discovery, The Biodesign Institute, Arizona State University, Tempe, AZ, 85287-7401, USA.
Ana Egatz-GomezSchool of Molecular Sciences, Arizona State University, Tempe, AZ, 85287-1604, USA.ORCID http://orcid.org/0000-0002-6080-943X
Sahir GandhiSchool of Molecular Sciences, Arizona State University, Tempe, AZ, 85287-1604, USA.
Gerrit BrehmInstitute for X-Ray Physics, University of Göttingen, Friedrich-Hund-Platz 1, 37077, Göttingen, Germany.
Chelsie E ConradSchool of Molecular Sciences, Arizona State University, Tempe, AZ, 85287-1604, USA.
Debra T HansenCenter for Applied Structural Discovery, The Biodesign Institute, Arizona State University, Tempe, AZ, 85287-7401, USA.ORCID http://orcid.org/0000-0003-4395-8297
Caleb MadsenCenter for Applied Structural Discovery, The Biodesign Institute, Arizona State University, Tempe, AZ, 85287-7401, USA.
Saša BajtDeutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.ORCID http://orcid.org/0000-0002-7163-7602
J Domingo Meza-AguilarEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Dominik OberthürCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.ORCID http://orcid.org/0000-0002-0894-9590
Max O WiedornCentre for Ultrafast Imaging, Universität Hamburg, Luruper Chaussee 149, 22761, Hamburg, Germany.
Holger FleckensteinCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.
Derek MendezCenter for Applied Structural Discovery, The Biodesign Institute, Arizona State University, Tempe, AZ, 85287-7401, USA.
Juraj KnoškaCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.ORCID http://orcid.org/0000-0001-7258-6256
Jose M Martin-GarciaSchool of Molecular Sciences, Arizona State University, Tempe, AZ, 85287-1604, USA.ORCID http://orcid.org/0000-0002-4558-3858
Hao HuCenter for Applied Structural Discovery, The Biodesign Institute, Arizona State University, Tempe, AZ, 85287-7401, USA.
Stella LisovaCenter for Applied Structural Discovery, The Biodesign Institute, Arizona State University, Tempe, AZ, 85287-7401, USA.ORCID http://orcid.org/0000-0002-6621-4285
Aschkan AllahgholiCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.
Yaroslav GevorkovCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.ORCID http://orcid.org/0000-0001-9273-7648
Kartik AyyerCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.ORCID http://orcid.org/0000-0002-6881-564X
Steve AplinCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.
Helen Mary GinnDivision of Structural Biology, University of Oxford, Oxford, OX1 2JD, United Kingdom.ORCID http://orcid.org/0000-0002-7894-4939
Heinz GraafsmaCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.
Andrew J MorganCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.
Dominic GreiffenbergPaul Scherrer Institute, Forschungsstrasse 111, 5232, Villigen, Switzerland.
Alexander KlujevCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.ORCID http://orcid.org/0000-0002-7490-2889
Torsten LaurusCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.
Jennifer PoehlsenCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.ORCID http://orcid.org/0000-0002-3114-8572
Ulrich TrunkCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.ORCID http://orcid.org/0000-0001-7951-4183
Davide MezzaPaul Scherrer Institute, Forschungsstrasse 111, 5232, Villigen, Switzerland.ORCID http://orcid.org/0000-0003-2439-6348
Bernd SchmidtPaul Scherrer Institute, Forschungsstrasse 111, 5232, Villigen, Switzerland.ORCID http://orcid.org/0000-0002-5778-0680
Manuela KuhnCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.
Raimund FrommeSchool of Molecular Sciences, Arizona State University, Tempe, AZ, 85287-1604, USA.ORCID http://orcid.org/0000-0003-4835-1080
Jolanta Sztuk-DambietzEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Natascha RaabEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Steffen HaufEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Alessandro SilenziEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Thomas MichelatEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.ORCID http://orcid.org/0000-0002-5689-8759
Chen XuEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Cyril DanilevskiEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Andrea ParentiEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Leonce MekindaEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Britta WeinhausenEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Grant MillsEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Patrik VagovicEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Yoonhee KimEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Henry KirkwoodEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.ORCID http://orcid.org/0000-0001-6792-3844
Richard BeanEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Johan BieleckiEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Stephan SternEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Klaus GiewekemeyerEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Adam R RoundEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Joachim SchulzEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.ORCID http://orcid.org/0000-0002-9310-8832
Katerina DörnerEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Thomas D GrantDepartment of Structural Biology, Jacobs School of Medicine and Biomedical Sciences, SUNY University at Buffalo, 955 Main St, Buffalo, NY, 14203, USA.ORCID http://orcid.org/0000-0002-8583-3700
Valerio MarianiCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.
Anton BartyCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Notkestrasse 85, 22607, Hamburg, Germany.
Adrian P MancusoEuropean XFEL, Holzkoppel 4, 22869, Schenefeld, Germany.
Uwe WeierstallCenter for Applied Structural Discovery, The Biodesign Institute, Arizona State University, Tempe, AZ, 85287-7401, USA.
John C H SpenceCenter for Applied Structural Discovery, The Biodesign Institute, Arizona State University, Tempe, AZ, 85287-7401, USA.
Henry N ChapmanCentre for Ultrafast Imaging, Universität Hamburg, Luruper Chaussee 149, 22761, Hamburg, Germany.ORCID http://orcid.org/0000-0002-4655-1743
Nadia ZatsepinCenter for Applied Structural Discovery, The Biodesign Institute, Arizona State University, Tempe, AZ, 85287-7401, USA.ORCID http://orcid.org/0000-0003-1757-0205
Petra FrommeSchool of Molecular Sciences, Arizona State University, Tempe, AZ, 85287-1604, USA.
Richard A KirianCenter for Applied Structural Discovery, The Biodesign Institute, Arizona State University, Tempe, AZ, 85287-7401, USA.
Alexandra RosSchool of Molecular Sciences, Arizona State University, Tempe, AZ, 85287-1604, USA. Alexandra.Ros@asu.edu.ORCID http://orcid.org/0000-0001-7709-8331
Funding
Femtosecond nano-crystallography of membrane proteinsR01GM095583 · NIGMS · ARIZONA STATE UNIVERSITY-TEMPE CAMPUS · PI FROMME, PETRA · 2010 to 2023
$4.3M3D NanoprinterS10OD021816 · OD · ARIZONA STATE UNIVERSITY-TEMPE CAMPUS · PI SPENCE, JOHN CHARLES · 2016 to 2016
$659kNIGMS NIH HHS R01 GM095583NIH HHS S10 OD021816
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.
Indexed as
ElectronsLab-On-A-Chip DevicesLasersAldehyde-LyasesCrystallographyEscherichia coli ProteinsHydrodynamics2-dehydro-3-deoxyphosphooctonate aldolaseAldehyde-LyasesEscherichia coli Proteins
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