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

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

doi:10.1038/s41467-020-18156-7

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

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

Research output: Contribution to journal › Article › peer-review

27 Scopus citations

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.

Original language	English (US)
Article number	4511
Journal	Nature communications
Volume	11
Issue number	1
DOIs	https://doi.org/10.1038/s41467-020-18156-7
State	Published - Dec 1 2020

ASJC Scopus subject areas

General Chemistry
General Biochemistry, Genetics and Molecular Biology
General Physics and Astronomy

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10.1038/s41467-020-18156-7

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Echelmeier, A., Cruz Villarreal, J., Messerschmidt, M., Kim, D., Coe, J. D., Thifault, D., Botha, S., Egatz-Gomez, A., Gandhi, S., Brehm, G., Conrad, C. E., Hansen, D. T., Madsen, C., Bajt, S., Meza-Aguilar, J. D., Oberthür, D., Wiedorn, M. O., Fleckenstein, H., Mendez, D., ... Ros, A. (2020). Segmented flow generator for serial crystallography at the European X-ray free electron laser. Nature communications, 11(1), Article 4511. https://doi.org/10.1038/s41467-020-18156-7

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 & Ros, A 2020, 'Segmented flow generator for serial crystallography at the European X-ray free electron laser', Nature communications, vol. 11, no. 1, 4511. https://doi.org/10.1038/s41467-020-18156-7

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title = "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.",

author = "Austin Echelmeier and {Cruz Villarreal}, Jorvani and Marc Messerschmidt and Daihyun Kim and Coe, {Jesse D.} and Darren Thifault and Sabine Botha and Ana Egatz-Gomez and Sahir Gandhi and Gerrit Brehm and Conrad, {Chelsie E.} and Hansen, {Debra T.} and Caleb Madsen and Sa{\v s}a Bajt and Meza-Aguilar, {J. Domingo} and Dominik Oberth{\"u}r and Wiedorn, {Max O.} and Holger Fleckenstein and Derek Mendez and Juraj Kno{\v s}ka and Martin-Garcia, {Jose M.} and Hao Hu and Stella Lisova and Aschkan Allahgholi and Yaroslav Gevorkov and Kartik Ayyer and Steve Aplin and Ginn, {Helen Mary} and Heinz Graafsma and Morgan, {Andrew J.} and Dominic Greiffenberg and Alexander Klujev and Torsten Laurus and Jennifer Poehlsen and Ulrich Trunk and Davide Mezza and Bernd Schmidt and Manuela Kuhn and Raimund Fromme and Jolanta Sztuk-Dambietz and Natascha Raab and Steffen Hauf and Alessandro Silenzi and Thomas Michelat and Chen Xu and Cyril Danilevski and Andrea Parenti and Leonce Mekinda and Britta Weinhausen and Grant Mills and Patrik Vagovic and Yoonhee Kim and Henry Kirkwood and Richard Bean and Johan Bielecki and Stephan Stern and Klaus Giewekemeyer and Round, {Adam R.} and Joachim Schulz and Katerina D{\"o}rner and Grant, {Thomas D.} and Valerio Mariani and Anton Barty and Mancuso, {Adrian P.} and Uwe Weierstall and Spence, {John C.H.} and Chapman, {Henry N.} and Nadia Zatsepin and Petra Fromme and Kirian, {Richard A.} and Alexandra Ros",

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AU - Echelmeier, Austin

AU - Cruz Villarreal, Jorvani

AU - Messerschmidt, Marc

AU - Kim, Daihyun

AU - Coe, Jesse D.

AU - Thifault, Darren

AU - Botha, Sabine

AU - Egatz-Gomez, Ana

AU - Gandhi, Sahir

AU - Brehm, Gerrit

AU - Conrad, Chelsie E.

AU - Hansen, Debra T.

AU - Madsen, Caleb

AU - Bajt, Saša

AU - Meza-Aguilar, J. Domingo

AU - Oberthür, Dominik

AU - Wiedorn, Max O.

AU - Fleckenstein, Holger

AU - Mendez, Derek

AU - Knoška, Juraj

AU - Martin-Garcia, Jose M.

AU - Hu, Hao

AU - Lisova, Stella

AU - Allahgholi, Aschkan

AU - Gevorkov, Yaroslav

AU - Ayyer, Kartik

AU - Aplin, Steve

AU - Ginn, Helen Mary

AU - Graafsma, Heinz

AU - Morgan, Andrew J.

AU - Greiffenberg, Dominic

AU - Klujev, Alexander

AU - Laurus, Torsten

AU - Poehlsen, Jennifer

AU - Trunk, Ulrich

AU - Mezza, Davide

AU - Schmidt, Bernd

AU - Kuhn, Manuela

AU - Fromme, Raimund

AU - Sztuk-Dambietz, Jolanta

AU - Raab, Natascha

AU - Hauf, Steffen

AU - Silenzi, Alessandro

AU - Michelat, Thomas

AU - Xu, Chen

AU - Danilevski, Cyril

AU - Parenti, Andrea

AU - Mekinda, Leonce

AU - Weinhausen, Britta

AU - Mills, Grant

AU - Vagovic, Patrik

AU - Kim, Yoonhee

AU - Kirkwood, Henry

AU - Bean, Richard

AU - Bielecki, Johan

AU - Stern, Stephan

AU - Giewekemeyer, Klaus

AU - Round, Adam R.

AU - Schulz, Joachim

AU - Dörner, Katerina

AU - Grant, Thomas D.

AU - Mariani, Valerio

AU - Barty, Anton

AU - Mancuso, Adrian P.

AU - Weierstall, Uwe

AU - Spence, John C.H.

AU - Chapman, Henry N.

AU - Zatsepin, Nadia

AU - Fromme, Petra

AU - Kirian, Richard A.

AU - Ros, Alexandra

PY - 2020/12/1

Y1 - 2020/12/1

N2 - 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.

AB - 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.

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

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KDO8PS Structure Determined at the EuXFEL using Segmented Flow Injection

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

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KDO8PS Structure Determined at the EuXFEL using Segmented Flow Injection

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