Coherent Diffraction Imaging(CDI)is an experimental technique to image isolated structures by recording the scattered light.The sample density can be recovered from the scattered field through a Fourier Transform oper...Coherent Diffraction Imaging(CDI)is an experimental technique to image isolated structures by recording the scattered light.The sample density can be recovered from the scattered field through a Fourier Transform operation.However,the phase of the field is lost during the measurement and has to be algorithmically retrieved.Here we present SPRING,an analysis framework tailored to X-ray Free Electron Laser(XFEL)single-shot single-particle diffraction data that implements the Memetic Phase Retrieval method to mitigate the shortcomings of conventional algorithms.We benchmark the approach on data acquired in two experimental campaigns at SwissFEL and European XFEL.Results reveal unprecedented stability and resilience of the algorithm’s behavior on the input parameters,and the capability of identifying the solution in conditions hardly treatable with conventional methods.A user-friendly implementation of SPRING is released as open-source software,aiming at being a reference tool for the CDI community at XFEL and synchrotron facilities.展开更多
The idea of using ultrashort X-ray pulses to obtain images of single proteins frozen in time has fascinated and inspired many.It was one of the arguments for building X-ray free-electron lasers.According to theory,the...The idea of using ultrashort X-ray pulses to obtain images of single proteins frozen in time has fascinated and inspired many.It was one of the arguments for building X-ray free-electron lasers.According to theory,the extremely intense pulses provide sufficient signal to dispense with using crystals as an amplifier,and the ultrashort pulse duration permits capturing the diffraction data before the sample inevitably explodes.This was first demonstrated on biological samples a decade ago on the giant mimivirus.Since then,a large collaboration has been pushing the limit of the smallest sample that can be imaged.The ability to capture snapshots on the timescale of atomic vibrations,while keeping the sample at room temperature,may allow probing the entire conformational phase space of macromolecules.Here we show the first observation of an X-ray diffraction pattern from a single protein,that of Escherichia coli GroEL which at 14 nm in diameter is the smallest biological sample ever imaged by X-rays,and demonstrate that the concept of diffraction before destruction extends to single proteins.From the pattern,it is possible to determine the approximate orientation of the protein.Our experiment demonstrates the feasibility of ultrafast imaging of single proteins,opening the way to single-molecule time-resolved studies on the femtosecond timescale.展开更多
基金the Swiss National Science Foundation (via grant no. 200021E_193642, grant no. 200021-232306, and the NCCR MUST)ETH Zurich (via collaborative grant 23-2ETH-050) for financial support+7 种基金MP, OV, and MB further acknowledge the Research Council of Finland for financial support (including projects 326291, 330118, and 341288)TF acknowledges funding by the Deutsche Forschungsgemeinschaft within CRC 1477 “Light-Matter Interactions at Interfaces” (project number 441234705)PHWS acknowledges support from the Swedish Research Council through project 2018-00740FRNCM acknowledges the Swedish Research Council (2018-00234 and 2019-06092) and the Carl Tryggers Stiftelse för Vetenskaplig Forskning (CTS 19-227)JAS acknowledges the Swedish Research Council (2023-06350)the Göran Gustafsson Foundation (2044)the Carl Tryggers Stiftelse för Vetenskaplig Forskning (CTS 21-1427)The Maloja instrument received funding from the Swiss National Science Foundation through R’Equip Grant No. 206021_182988. We thank the IT Services Group (ISG) of the Department of Physics at ETH Zurich for the excellent support and management of the computing hardware on which the spring software has been developed and tested.
文摘Coherent Diffraction Imaging(CDI)is an experimental technique to image isolated structures by recording the scattered light.The sample density can be recovered from the scattered field through a Fourier Transform operation.However,the phase of the field is lost during the measurement and has to be algorithmically retrieved.Here we present SPRING,an analysis framework tailored to X-ray Free Electron Laser(XFEL)single-shot single-particle diffraction data that implements the Memetic Phase Retrieval method to mitigate the shortcomings of conventional algorithms.We benchmark the approach on data acquired in two experimental campaigns at SwissFEL and European XFEL.Results reveal unprecedented stability and resilience of the algorithm’s behavior on the input parameters,and the capability of identifying the solution in conditions hardly treatable with conventional methods.A user-friendly implementation of SPRING is released as open-source software,aiming at being a reference tool for the CDI community at XFEL and synchrotron facilities.
基金supported by the Universität Hamburg and DFG grant numbers(INST 152/772-1|152/774-1|152/775-1|152/776-1|152/777-1 FUGG)We acknowledge the support of funding from:Cluster of Excellence‘CUI:Advanced Imaging of Matter’of the Deutsche Forschungsgemeinschaft(DFG)-EXC 2056-project ID 390715994+7 种基金ERC-2013-CoG COMOTION 614507NFR 240770Fellowship from the Joachim Herz Stiftung(P.L.X.)P.L.X.and H.N.C.acknowledge support from the Human Frontiers Science Program(RGP0010/2017)J.H.acknowledges support from the European Development Fund:Structural dynamics of biomolecular systems(ELIBIO)(CZ.02.1.01/0.0/0.0/15_003/0000447)EMBO long-term fellowship(ALTF 356-2018)awarded to L.E.F.the Röntgen-Ångström Cluster(2015-06107 and 2019-06092)the Swedish Research Council(2017-05336,2018-00234 and 2019-03935)the Swedish Foundation for Strategic Research(ITM17-0455).
文摘The idea of using ultrashort X-ray pulses to obtain images of single proteins frozen in time has fascinated and inspired many.It was one of the arguments for building X-ray free-electron lasers.According to theory,the extremely intense pulses provide sufficient signal to dispense with using crystals as an amplifier,and the ultrashort pulse duration permits capturing the diffraction data before the sample inevitably explodes.This was first demonstrated on biological samples a decade ago on the giant mimivirus.Since then,a large collaboration has been pushing the limit of the smallest sample that can be imaged.The ability to capture snapshots on the timescale of atomic vibrations,while keeping the sample at room temperature,may allow probing the entire conformational phase space of macromolecules.Here we show the first observation of an X-ray diffraction pattern from a single protein,that of Escherichia coli GroEL which at 14 nm in diameter is the smallest biological sample ever imaged by X-rays,and demonstrate that the concept of diffraction before destruction extends to single proteins.From the pattern,it is possible to determine the approximate orientation of the protein.Our experiment demonstrates the feasibility of ultrafast imaging of single proteins,opening the way to single-molecule time-resolved studies on the femtosecond timescale.
