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


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.

SUBMITTER: Echelmeier A 

PROVIDER: S-EPMC7481229 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature

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

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

Nature communications 20200909 1


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 s  ...[more]

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