1. Segmented flow generator for serial crystallography at the European X-ray free electron laser.
- Author
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Echelmeier A, Cruz Villarreal J, Messerschmidt M, Kim D, Coe JD, Thifault D, Botha S, Egatz-Gomez A, Gandhi S, Brehm G, Conrad CE, Hansen DT, Madsen C, Bajt S, Meza-Aguilar JD, Oberthür D, Wiedorn MO, Fleckenstein H, Mendez D, Knoška J, Martin-Garcia JM, Hu H, Lisova S, Allahgholi A, Gevorkov Y, Ayyer K, Aplin S, Ginn HM, Graafsma H, Morgan AJ, Greiffenberg D, Klujev A, Laurus T, Poehlsen J, Trunk U, Mezza D, Schmidt B, Kuhn M, Fromme R, Sztuk-Dambietz J, Raab N, Hauf S, Silenzi A, Michelat T, Xu C, Danilevski C, Parenti A, Mekinda L, Weinhausen B, Mills G, Vagovic P, Kim Y, Kirkwood H, Bean R, Bielecki J, Stern S, Giewekemeyer K, Round AR, Schulz J, Dörner K, Grant TD, Mariani V, Barty A, Mancuso AP, Weierstall U, Spence JCH, Chapman HN, Zatsepin N, Fromme P, Kirian RA, and Ros A
- Subjects
- Aldehyde-Lyases ultrastructure, Escherichia coli Proteins ultrastructure, Hydrodynamics, Crystallography instrumentation, Electrons, Lab-On-A-Chip Devices, Lasers
- 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.
- Published
- 2020
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