Megahertz pulse trains enable multi-hit serial femtosecond crystallography experiments at X-ray free electron lasers

Susannah Holmes; Henry J. Kirkwood; Richard Bean; Klaus Giewekemeyer; Andrew V. Martin; Marjan Hadian-Jazi; Max O. Wiedorn; Dominik Oberthür; Hugh Marman; Luigi Adriano; Nasser Al-Qudami; Saša Bajt; Imrich Barák; Sadia Bari; Johan Bielecki; Sandor Brockhauser; Mathew A. Coleman; Francisco Cruz-Mazo; Cyril Danilevski; Katerina Dörner; Alfonso M. Gañán-Calvo; Rita Graceffa; Hans Fanghor; Michael Heymann; Matthias Frank; Alexander Kaukher; Yoonhee Kim; Bostjan Kobe; Juraj Knoška; Torsten Laurus; Romain Letrun; Luis Maia; Marc Messerschmidt; Markus Metz; Thomas Michelat; Grant Mills; Serguei Molodtsov; Diana C.F. Monteiro; Andrew J. Morgan; Astrid Münnich; Gisel E. Peña Murillo; Gianpietro Previtali; Adam Round; Tokushi Sato; Robin Schubert; Joachim Schulz; Megan Shelby; Carolin Seuring; Jonas A. Sellberg; Marcin Sikorski; Alessandro Silenzi; Stephan Stern; Jola Sztuk-Dambietz; Janusz Szuba; Martin Trebbin; Patrick Vagovic; Thomas Ve; Britta Weinhausen; Krzysztof Wrona; Paul Lourdu Xavier; Chen Xu; Oleksandr Yefanov; Keith A. Nugent; Henry N. Chapman; Adrian P. Mancuso; Anton Barty; Brian Abbey; Connie Darmanin

doi:10.1038/s41467-022-32434-6

Megahertz pulse trains enable multi-hit serial femtosecond crystallography experiments at X-ray free electron lasers

Susannah Holmes, Henry J. Kirkwood, Richard Bean, Klaus Giewekemeyer, Andrew V. Martin, Marjan Hadian-Jazi, Max O. Wiedorn, Dominik Oberthür, Hugh Marman, Luigi Adriano, Nasser Al-Qudami, Saša Bajt, Imrich Barák, Sadia Bari, Johan Bielecki, Sandor Brockhauser, Mathew A. Coleman, Francisco Cruz-Mazo, Cyril Danilevski, Katerina DörnerAlfonso M. Gañán-Calvo, Rita Graceffa, Hans Fanghor, Michael Heymann, Matthias Frank, Alexander Kaukher, Yoonhee Kim, Bostjan Kobe, Juraj Knoška, Torsten Laurus, Romain Letrun, Luis Maia, Marc Messerschmidt, Markus Metz, Thomas Michelat, Grant Mills, Serguei Molodtsov, Diana C.F. Monteiro, Andrew J. Morgan, Astrid Münnich, Gisel E. Peña Murillo, Gianpietro Previtali, Adam Round, Tokushi Sato, Robin Schubert, Joachim Schulz, Megan Shelby, Carolin Seuring, Jonas A. Sellberg, Marcin Sikorski, Alessandro Silenzi, Stephan Stern, Jola Sztuk-Dambietz, Janusz Szuba, Martin Trebbin, Patrick Vagovic, Thomas Ve, Britta Weinhausen, Krzysztof Wrona, Paul Lourdu Xavier, Chen Xu, Oleksandr Yefanov, Keith A. Nugent, Henry N. Chapman, Adrian P. Mancuso, Anton Barty, Brian Abbey, Connie Darmanin

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

1 Scopus citations

Abstract

The European X-ray Free Electron Laser (XFEL) and Linac Coherent Light Source (LCLS) II are extremely intense sources of X-rays capable of generating Serial Femtosecond Crystallography (SFX) data at megahertz (MHz) repetition rates. Previous work has shown that it is possible to use consecutive X-ray pulses to collect diffraction patterns from individual crystals. Here, we exploit the MHz pulse structure of the European XFEL to obtain two complete datasets from the same lysozyme crystal, first hit and the second hit, before it exits the beam. The two datasets, separated by <1 µs, yield up to 2.1 Å resolution structures. Comparisons between the two structures reveal no indications of radiation damage or significant changes within the active site, consistent with the calculated dose estimates. This demonstrates MHz SFX can be used as a tool for tracking sub-microsecond structural changes in individual single crystals, a technique we refer to as multi-hit SFX.

Original language	English (US)
Article number	4708
Journal	Nature communications
Volume	13
Issue number	1
DOIs	https://doi.org/10.1038/s41467-022-32434-6
State	Published - Dec 2022
Externally published	Yes

ASJC Scopus subject areas

Chemistry(all)
Biochemistry, Genetics and Molecular Biology(all)
General
Physics and Astronomy(all)

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10.1038/s41467-022-32434-6

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Holmes, S., Kirkwood, H. J., Bean, R., Giewekemeyer, K., Martin, A. V., Hadian-Jazi, M., Wiedorn, M. O., Oberthür, D., Marman, H., Adriano, L., Al-Qudami, N., Bajt, S., Barák, I., Bari, S., Bielecki, J., Brockhauser, S., Coleman, M. A., Cruz-Mazo, F., Danilevski, C., ... Darmanin, C. (2022). Megahertz pulse trains enable multi-hit serial femtosecond crystallography experiments at X-ray free electron lasers. Nature communications, 13(1), [4708]. https://doi.org/10.1038/s41467-022-32434-6

Holmes, S, Kirkwood, HJ, Bean, R, Giewekemeyer, K, Martin, AV, Hadian-Jazi, M, Wiedorn, MO, Oberthür, D, Marman, H, Adriano, L, Al-Qudami, N, Bajt, S, Barák, I, Bari, S, Bielecki, J, Brockhauser, S, Coleman, MA, Cruz-Mazo, F, Danilevski, C, Dörner, K, Gañán-Calvo, AM, Graceffa, R, Fanghor, H, Heymann, M, Frank, M, Kaukher, A, Kim, Y, Kobe, B, Knoška, J, Laurus, T, Letrun, R, Maia, L, Messerschmidt, M, Metz, M, Michelat, T, Mills, G, Molodtsov, S, Monteiro, DCF, Morgan, AJ, Münnich, A, Peña Murillo, GE, Previtali, G, Round, A, Sato, T, Schubert, R, Schulz, J, Shelby, M, Seuring, C, Sellberg, JA, Sikorski, M, Silenzi, A, Stern, S, Sztuk-Dambietz, J, Szuba, J, Trebbin, M, Vagovic, P, Ve, T, Weinhausen, B, Wrona, K, Xavier, PL, Xu, C, Yefanov, O, Nugent, KA, Chapman, HN, Mancuso, AP, Barty, A, Abbey, B & Darmanin, C 2022, 'Megahertz pulse trains enable multi-hit serial femtosecond crystallography experiments at X-ray free electron lasers', Nature communications, vol. 13, no. 1, 4708. https://doi.org/10.1038/s41467-022-32434-6

@article{de3af8d170e84950b2d5fdb79ab59c25,

title = "Megahertz pulse trains enable multi-hit serial femtosecond crystallography experiments at X-ray free electron lasers",

abstract = "The European X-ray Free Electron Laser (XFEL) and Linac Coherent Light Source (LCLS) II are extremely intense sources of X-rays capable of generating Serial Femtosecond Crystallography (SFX) data at megahertz (MHz) repetition rates. Previous work has shown that it is possible to use consecutive X-ray pulses to collect diffraction patterns from individual crystals. Here, we exploit the MHz pulse structure of the European XFEL to obtain two complete datasets from the same lysozyme crystal, first hit and the second hit, before it exits the beam. The two datasets, separated by <1 µs, yield up to 2.1 {\AA} resolution structures. Comparisons between the two structures reveal no indications of radiation damage or significant changes within the active site, consistent with the calculated dose estimates. This demonstrates MHz SFX can be used as a tool for tracking sub-microsecond structural changes in individual single crystals, a technique we refer to as multi-hit SFX.",

author = "Susannah Holmes and Kirkwood, {Henry J.} and Richard Bean and Klaus Giewekemeyer and Martin, {Andrew V.} and Marjan Hadian-Jazi and Wiedorn, {Max O.} and Dominik Oberth{\"u}r and Hugh Marman and Luigi Adriano and Nasser Al-Qudami and Sa{\v s}a Bajt and Imrich Bar{\'a}k and Sadia Bari and Johan Bielecki and Sandor Brockhauser and Coleman, {Mathew A.} and Francisco Cruz-Mazo and Cyril Danilevski and Katerina D{\"o}rner and Ga{\~n}{\'a}n-Calvo, {Alfonso M.} and Rita Graceffa and Hans Fanghor and Michael Heymann and Matthias Frank and Alexander Kaukher and Yoonhee Kim and Bostjan Kobe and Juraj Kno{\v s}ka and Torsten Laurus and Romain Letrun and Luis Maia and Marc Messerschmidt and Markus Metz and Thomas Michelat and Grant Mills and Serguei Molodtsov and Monteiro, {Diana C.F.} and Morgan, {Andrew J.} and Astrid M{\"u}nnich and {Pe{\~n}a Murillo}, {Gisel E.} and Gianpietro Previtali and Adam Round and Tokushi Sato and Robin Schubert and Joachim Schulz and Megan Shelby and Carolin Seuring and Sellberg, {Jonas A.} and Marcin Sikorski and Alessandro Silenzi and Stephan Stern and Jola Sztuk-Dambietz and Janusz Szuba and Martin Trebbin and Patrick Vagovic and Thomas Ve and Britta Weinhausen and Krzysztof Wrona and Xavier, {Paul Lourdu} and Chen Xu and Oleksandr Yefanov and Nugent, {Keith A.} and Chapman, {Henry N.} and Mancuso, {Adrian P.} and Anton Barty and Brian Abbey and Connie Darmanin",

note = "Funding Information: We acknowledge European XFEL in Schenefeld, Germany, for provision of X-ray free electron laser beam-time at Scientific Instrument SPB/SFX and use of the XBI biological sample preparation laboratory, enabled by the XBI User Consortium and would like to thank the instrument group and facility staff for their assistance and everyone who attended the first experiment. We would also like to acknowledge the funding support for B.A., C.D, K.A.N, M. H-J and S.H from Australian Research Council Centre of Excellence in Advanced Molecular Imaging (CE140100011), www.imagingcoe.org and the Australian Nuclear Science and Technology Organisation (ANSTO); the International Synchrotron Access Program (ISAP) managed by the Australian Synchrotron, part of ANSTO, and funded by the Australian Government. M.L.S., M.A.C. and M.F. performed, in part, under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344. M.L.S., M.A.C. and M.F. were supported by the NIH grant 1R01GM117342-01. I.B acknowledges support by Ministry of Education, Science, Research and Sport of the Slovak Republic and by grant from the Slovak Research and Development Agency under contract APVV-14-0181. M.L.S., M.A.C. and M.F. performed, in part, under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344. M.L.S., M.A.C. and M.F. were supported by the NIH grant 1R01GM117342-01. T.V is a recipient of ARC DECRA (DE170100783). B.K work is funded by National Health and Medical Research Council (NHMRC) grants 1107804 and 1160570 to BK and TV; BK is Australian Research Council (ARC) Laureate Fellow (FL180100109). F.C-M and A.M.G-C acknowledge the Spanish Agencia Estatal de Investigaci{\'o}n (grant no. PID2019-108278RB), and the Junta de Andaluc{\'i}a (grant no. P18-FR-3375). S.Bari acknowledged the Helmholtz Initiative and Networking Fund through the Young Investigators Program and the Deutsche Forschungsgemeinschaft SFB755/B03. PLX acknowledges a fellowship from the Joachim Herz Stiftung. Additional support was provided by the Cluster of Excellence {\textquoteleft}CUI: Advanced Imaging of Matter{\textquoteright} of the Deutsche Forschungsgemeinschaft (DFG) – EXC 2056 – project ID 390715994 and by the Human Frontiers Science Program (RGP0010/2017). Funding Information: We acknowledge European XFEL in Schenefeld, Germany, for provision of X-ray free electron laser beam-time at Scientific Instrument SPB/SFX and use of the XBI biological sample preparation laboratory, enabled by the XBI User Consortium and would like to thank the instrument group and facility staff for their assistance and everyone who attended the first experiment. We would also like to acknowledge the funding support for B.A., C.D, K.A.N, M. H-J and S.H from Australian Research Council Centre of Excellence in Advanced Molecular Imaging (CE140100011), www.imagingcoe.org and the Australian Nuclear Science and Technology Organisation (ANSTO); the International Synchrotron Access Program (ISAP) managed by the Australian Synchrotron, part of ANSTO, and funded by the Australian Government. M.L.S., M.A.C. and M.F. performed, in part, under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344. M.L.S., M.A.C. and M.F. were supported by the NIH grant 1R01GM117342-01. I.B acknowledges support by Ministry of Education, Science, Research and Sport of the Slovak Republic and by grant from the Slovak Research and Development Agency under contract APVV-14-0181. M.L.S., M.A.C. and M.F. performed, in part, under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344. M.L.S., M.A.C. and M.F. were supported by the NIH grant 1R01GM117342-01. T.V is a recipient of ARC DECRA (DE170100783). B.K work is funded by National Health and Medical Research Council (NHMRC) grants 1107804 and 1160570 to BK and TV; BK is Australian Research Council (ARC) Laureate Fellow (FL180100109). F.C-M and A.M.G-C acknowledge the Spanish Agencia Estatal de Investigaci{\'o}n (grant no. PID2019-108278RB), and the Junta de Andaluc{\'i}a (grant no. P18-FR-3375). S.Bari acknowledged the Helmholtz Initiative and Networking Fund through the Young Investigators Program and the Deutsche Forschungsgemeinschaft SFB755/B03. PLX acknowledges a fellowship from the Joachim Herz Stiftung. Additional support was provided by the Cluster of Excellence {\textquoteleft}CUI: Advanced Imaging of Matter{\textquoteright} of the Deutsche Forschungsgemeinschaft (DFG) – EXC 2056 – project ID 390715994 and by the Human Frontiers Science Program (RGP0010/2017). Publisher Copyright: {\textcopyright} 2022, The Author(s).",

year = "2022",

month = dec,

doi = "10.1038/s41467-022-32434-6",

language = "English (US)",

volume = "13",

journal = "Nature Communications",

issn = "2041-1723",

publisher = "Nature Publishing Group",

number = "1",

}

TY - JOUR

T1 - Megahertz pulse trains enable multi-hit serial femtosecond crystallography experiments at X-ray free electron lasers

AU - Holmes, Susannah

AU - Kirkwood, Henry J.

AU - Bean, Richard

AU - Giewekemeyer, Klaus

AU - Martin, Andrew V.

AU - Hadian-Jazi, Marjan

AU - Wiedorn, Max O.

AU - Oberthür, Dominik

AU - Marman, Hugh

AU - Adriano, Luigi

AU - Al-Qudami, Nasser

AU - Bajt, Saša

AU - Barák, Imrich

AU - Bari, Sadia

AU - Bielecki, Johan

AU - Brockhauser, Sandor

AU - Coleman, Mathew A.

AU - Cruz-Mazo, Francisco

AU - Danilevski, Cyril

AU - Dörner, Katerina

AU - Gañán-Calvo, Alfonso M.

AU - Graceffa, Rita

AU - Fanghor, Hans

AU - Heymann, Michael

AU - Frank, Matthias

AU - Kaukher, Alexander

AU - Kim, Yoonhee

AU - Kobe, Bostjan

AU - Knoška, Juraj

AU - Laurus, Torsten

AU - Letrun, Romain

AU - Maia, Luis

AU - Messerschmidt, Marc

AU - Metz, Markus

AU - Michelat, Thomas

AU - Mills, Grant

AU - Molodtsov, Serguei

AU - Monteiro, Diana C.F.

AU - Morgan, Andrew J.

AU - Münnich, Astrid

AU - Peña Murillo, Gisel E.

AU - Previtali, Gianpietro

AU - Round, Adam

AU - Sato, Tokushi

AU - Schubert, Robin

AU - Schulz, Joachim

AU - Shelby, Megan

AU - Seuring, Carolin

AU - Sellberg, Jonas A.

AU - Sikorski, Marcin

AU - Silenzi, Alessandro

AU - Stern, Stephan

AU - Sztuk-Dambietz, Jola

AU - Szuba, Janusz

AU - Trebbin, Martin

AU - Vagovic, Patrick

AU - Ve, Thomas

AU - Weinhausen, Britta

AU - Wrona, Krzysztof

AU - Xavier, Paul Lourdu

AU - Xu, Chen

AU - Yefanov, Oleksandr

AU - Nugent, Keith A.

AU - Chapman, Henry N.

AU - Mancuso, Adrian P.

AU - Barty, Anton

AU - Abbey, Brian

AU - Darmanin, Connie

N1 - Funding Information: We acknowledge European XFEL in Schenefeld, Germany, for provision of X-ray free electron laser beam-time at Scientific Instrument SPB/SFX and use of the XBI biological sample preparation laboratory, enabled by the XBI User Consortium and would like to thank the instrument group and facility staff for their assistance and everyone who attended the first experiment. We would also like to acknowledge the funding support for B.A., C.D, K.A.N, M. H-J and S.H from Australian Research Council Centre of Excellence in Advanced Molecular Imaging (CE140100011), www.imagingcoe.org and the Australian Nuclear Science and Technology Organisation (ANSTO); the International Synchrotron Access Program (ISAP) managed by the Australian Synchrotron, part of ANSTO, and funded by the Australian Government. M.L.S., M.A.C. and M.F. performed, in part, under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344. M.L.S., M.A.C. and M.F. were supported by the NIH grant 1R01GM117342-01. I.B acknowledges support by Ministry of Education, Science, Research and Sport of the Slovak Republic and by grant from the Slovak Research and Development Agency under contract APVV-14-0181. M.L.S., M.A.C. and M.F. performed, in part, under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344. M.L.S., M.A.C. and M.F. were supported by the NIH grant 1R01GM117342-01. T.V is a recipient of ARC DECRA (DE170100783). B.K work is funded by National Health and Medical Research Council (NHMRC) grants 1107804 and 1160570 to BK and TV; BK is Australian Research Council (ARC) Laureate Fellow (FL180100109). F.C-M and A.M.G-C acknowledge the Spanish Agencia Estatal de Investigación (grant no. PID2019-108278RB), and the Junta de Andalucía (grant no. P18-FR-3375). S.Bari acknowledged the Helmholtz Initiative and Networking Fund through the Young Investigators Program and the Deutsche Forschungsgemeinschaft SFB755/B03. PLX acknowledges a fellowship from the Joachim Herz Stiftung. Additional support was provided by the Cluster of Excellence ‘CUI: Advanced Imaging of Matter’ of the Deutsche Forschungsgemeinschaft (DFG) – EXC 2056 – project ID 390715994 and by the Human Frontiers Science Program (RGP0010/2017). Funding Information: We acknowledge European XFEL in Schenefeld, Germany, for provision of X-ray free electron laser beam-time at Scientific Instrument SPB/SFX and use of the XBI biological sample preparation laboratory, enabled by the XBI User Consortium and would like to thank the instrument group and facility staff for their assistance and everyone who attended the first experiment. We would also like to acknowledge the funding support for B.A., C.D, K.A.N, M. H-J and S.H from Australian Research Council Centre of Excellence in Advanced Molecular Imaging (CE140100011), www.imagingcoe.org and the Australian Nuclear Science and Technology Organisation (ANSTO); the International Synchrotron Access Program (ISAP) managed by the Australian Synchrotron, part of ANSTO, and funded by the Australian Government. M.L.S., M.A.C. and M.F. performed, in part, under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344. M.L.S., M.A.C. and M.F. were supported by the NIH grant 1R01GM117342-01. I.B acknowledges support by Ministry of Education, Science, Research and Sport of the Slovak Republic and by grant from the Slovak Research and Development Agency under contract APVV-14-0181. M.L.S., M.A.C. and M.F. performed, in part, under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344. M.L.S., M.A.C. and M.F. were supported by the NIH grant 1R01GM117342-01. T.V is a recipient of ARC DECRA (DE170100783). B.K work is funded by National Health and Medical Research Council (NHMRC) grants 1107804 and 1160570 to BK and TV; BK is Australian Research Council (ARC) Laureate Fellow (FL180100109). F.C-M and A.M.G-C acknowledge the Spanish Agencia Estatal de Investigación (grant no. PID2019-108278RB), and the Junta de Andalucía (grant no. P18-FR-3375). S.Bari acknowledged the Helmholtz Initiative and Networking Fund through the Young Investigators Program and the Deutsche Forschungsgemeinschaft SFB755/B03. PLX acknowledges a fellowship from the Joachim Herz Stiftung. Additional support was provided by the Cluster of Excellence ‘CUI: Advanced Imaging of Matter’ of the Deutsche Forschungsgemeinschaft (DFG) – EXC 2056 – project ID 390715994 and by the Human Frontiers Science Program (RGP0010/2017). Publisher Copyright: © 2022, The Author(s).

PY - 2022/12

Y1 - 2022/12

N2 - The European X-ray Free Electron Laser (XFEL) and Linac Coherent Light Source (LCLS) II are extremely intense sources of X-rays capable of generating Serial Femtosecond Crystallography (SFX) data at megahertz (MHz) repetition rates. Previous work has shown that it is possible to use consecutive X-ray pulses to collect diffraction patterns from individual crystals. Here, we exploit the MHz pulse structure of the European XFEL to obtain two complete datasets from the same lysozyme crystal, first hit and the second hit, before it exits the beam. The two datasets, separated by <1 µs, yield up to 2.1 Å resolution structures. Comparisons between the two structures reveal no indications of radiation damage or significant changes within the active site, consistent with the calculated dose estimates. This demonstrates MHz SFX can be used as a tool for tracking sub-microsecond structural changes in individual single crystals, a technique we refer to as multi-hit SFX.

AB - The European X-ray Free Electron Laser (XFEL) and Linac Coherent Light Source (LCLS) II are extremely intense sources of X-rays capable of generating Serial Femtosecond Crystallography (SFX) data at megahertz (MHz) repetition rates. Previous work has shown that it is possible to use consecutive X-ray pulses to collect diffraction patterns from individual crystals. Here, we exploit the MHz pulse structure of the European XFEL to obtain two complete datasets from the same lysozyme crystal, first hit and the second hit, before it exits the beam. The two datasets, separated by <1 µs, yield up to 2.1 Å resolution structures. Comparisons between the two structures reveal no indications of radiation damage or significant changes within the active site, consistent with the calculated dose estimates. This demonstrates MHz SFX can be used as a tool for tracking sub-microsecond structural changes in individual single crystals, a technique we refer to as multi-hit SFX.

UR - http://www.scopus.com/inward/record.url?scp=85135812771&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85135812771&partnerID=8YFLogxK

U2 - 10.1038/s41467-022-32434-6

DO - 10.1038/s41467-022-32434-6

M3 - Article

C2 - 35953469

AN - SCOPUS:85135812771

VL - 13

JO - Nature Communications

JF - Nature Communications

SN - 2041-1723

IS - 1

M1 - 4708

ER -

Megahertz pulse trains enable multi-hit serial femtosecond crystallography experiments at X-ray free electron lasers

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