A fixed-target platform for serial femtosecond crystallography in a hydrated environment

For serial femtosecond crystallography at X-ray free-electron lasers, which entails collection of single-pulse diffraction patterns from a constantly refreshed supply of microcrystalline sample, delivery of the sample into the X-ray beam path while maintaining low background remains a technical chal...

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Main Authors: M. L. Shelby, D. Gilbile, T. D. Grant, C. Seuring, B. W. Segelke, W. He, A. C. Evans, T. Pakendorf, P. Fischer, M. S. Hunter, A. Batyuk, M. Barthelmess, A. Meents, M. A. Coleman, T. L. Kuhl, M. Frank
Format: Article
Language:English
Published: International Union of Crystallography 2020-01-01
Series:IUCrJ
Subjects:
Online Access:http://scripts.iucr.org/cgi-bin/paper?S2052252519014003
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spelling doaj-e421557afb46417b922f859317082c872020-11-25T02:16:32ZengInternational Union of CrystallographyIUCrJ2052-25252020-01-0171304110.1107/S2052252519014003ec5015A fixed-target platform for serial femtosecond crystallography in a hydrated environmentM. L. Shelby0D. Gilbile1T. D. Grant2C. Seuring3B. W. Segelke4W. He5A. C. Evans6T. Pakendorf7P. Fischer8M. S. Hunter9A. Batyuk10M. Barthelmess11A. Meents12M. A. Coleman13T. L. Kuhl14M. Frank15Lawrence Livermore National Laboratory, Livermore, CA 94550, USAUniversity of California at Davis, California, USADepartment of Structural Biology, Jacobs School of Medicine and Biomedical Sciences, Hauptman-Woodward Institute, SUNY University at Buffalo, Buffalo, New York, USACenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Hamburg, GermanyLawrence Livermore National Laboratory, Livermore, CA 94550, USALawrence Livermore National Laboratory, Livermore, CA 94550, USALawrence Livermore National Laboratory, Livermore, CA 94550, USACenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Hamburg, GermanyCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Hamburg, GermanyLinac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California, USALinac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California, USACenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Hamburg, GermanyCenter for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron, Hamburg, GermanyLawrence Livermore National Laboratory, Livermore, CA 94550, USAUniversity of California at Davis, California, USALawrence Livermore National Laboratory, Livermore, CA 94550, USAFor serial femtosecond crystallography at X-ray free-electron lasers, which entails collection of single-pulse diffraction patterns from a constantly refreshed supply of microcrystalline sample, delivery of the sample into the X-ray beam path while maintaining low background remains a technical challenge for some experiments, especially where this methodology is applied to relatively low-ordered samples or those difficult to purify and crystallize in large quantities. This work demonstrates a scheme to encapsulate biological samples using polymer thin films and graphene to maintain sample hydration in vacuum conditions. The encapsulated sample is delivered into the X-ray beam on fixed targets for rapid scanning using the Roadrunner fixed-target system towards a long-term goal of low-background measurements on weakly diffracting samples. As a proof of principle, we used microcrystals of the 24 kDa rapid encystment protein (REP24) to provide a benchmark for polymer/graphene sandwich performance. The REP24 microcrystal unit cell obtained from our sandwiched in-vacuum sample was consistent with previously established unit-cell parameters and with those measured by us without encapsulation in humidified helium, indicating that the platform is robust against evaporative losses. While significant scattering from water was observed because of the sample-deposition method, the polymer/graphene sandwich itself was shown to contribute minimally to background scattering.http://scripts.iucr.org/cgi-bin/paper?S2052252519014003serial crystallographysample deliveryxfelsin-vacuum studiesfixed-target platformsgraphenepolymersthin filmsmicrocrystalssample hydration
collection DOAJ
language English
format Article
sources DOAJ
author M. L. Shelby
D. Gilbile
T. D. Grant
C. Seuring
B. W. Segelke
W. He
A. C. Evans
T. Pakendorf
P. Fischer
M. S. Hunter
A. Batyuk
M. Barthelmess
A. Meents
M. A. Coleman
T. L. Kuhl
M. Frank
spellingShingle M. L. Shelby
D. Gilbile
T. D. Grant
C. Seuring
B. W. Segelke
W. He
A. C. Evans
T. Pakendorf
P. Fischer
M. S. Hunter
A. Batyuk
M. Barthelmess
A. Meents
M. A. Coleman
T. L. Kuhl
M. Frank
A fixed-target platform for serial femtosecond crystallography in a hydrated environment
IUCrJ
serial crystallography
sample delivery
xfels
in-vacuum studies
fixed-target platforms
graphene
polymers
thin films
microcrystals
sample hydration
author_facet M. L. Shelby
D. Gilbile
T. D. Grant
C. Seuring
B. W. Segelke
W. He
A. C. Evans
T. Pakendorf
P. Fischer
M. S. Hunter
A. Batyuk
M. Barthelmess
A. Meents
M. A. Coleman
T. L. Kuhl
M. Frank
author_sort M. L. Shelby
title A fixed-target platform for serial femtosecond crystallography in a hydrated environment
title_short A fixed-target platform for serial femtosecond crystallography in a hydrated environment
title_full A fixed-target platform for serial femtosecond crystallography in a hydrated environment
title_fullStr A fixed-target platform for serial femtosecond crystallography in a hydrated environment
title_full_unstemmed A fixed-target platform for serial femtosecond crystallography in a hydrated environment
title_sort fixed-target platform for serial femtosecond crystallography in a hydrated environment
publisher International Union of Crystallography
series IUCrJ
issn 2052-2525
publishDate 2020-01-01
description For serial femtosecond crystallography at X-ray free-electron lasers, which entails collection of single-pulse diffraction patterns from a constantly refreshed supply of microcrystalline sample, delivery of the sample into the X-ray beam path while maintaining low background remains a technical challenge for some experiments, especially where this methodology is applied to relatively low-ordered samples or those difficult to purify and crystallize in large quantities. This work demonstrates a scheme to encapsulate biological samples using polymer thin films and graphene to maintain sample hydration in vacuum conditions. The encapsulated sample is delivered into the X-ray beam on fixed targets for rapid scanning using the Roadrunner fixed-target system towards a long-term goal of low-background measurements on weakly diffracting samples. As a proof of principle, we used microcrystals of the 24 kDa rapid encystment protein (REP24) to provide a benchmark for polymer/graphene sandwich performance. The REP24 microcrystal unit cell obtained from our sandwiched in-vacuum sample was consistent with previously established unit-cell parameters and with those measured by us without encapsulation in humidified helium, indicating that the platform is robust against evaporative losses. While significant scattering from water was observed because of the sample-deposition method, the polymer/graphene sandwich itself was shown to contribute minimally to background scattering.
topic serial crystallography
sample delivery
xfels
in-vacuum studies
fixed-target platforms
graphene
polymers
thin films
microcrystals
sample hydration
url http://scripts.iucr.org/cgi-bin/paper?S2052252519014003
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