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

Abstract

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 backgroundmore » scattering.« less

Authors:
; ; ORCiD logo; ; ; ; ; ; ; ; ; ; ORCiD logo; ; ;
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1605168
Grant/Contract Number:  
[AC52-07NA27344; AC02-76SF00515; 1R01GM117342-01; STC-1231306]
Resource Type:
Accepted Manuscript
Journal Name:
IUCrJ
Additional Journal Information:
[ Journal Volume: 7; Journal Issue: 1]; Journal ID: ISSN 2052-2525
Country of Publication:
United States
Language:
English

Citation Formats

Shelby, M. L., Gilbile, D., Grant, T. D., Seuring, C., Segelke, B. W., He, W., Evans, A. C., Pakendorf, T., Fischer, P., Hunter, M. S., Batyuk, A., Barthelmess, M., Meents, A., Coleman, M. A., Kuhl, T. L., and Frank, M. A fixed-target platform for serial femtosecond crystallography in a hydrated environment. United States: N. p., 2020. Web. doi:10.1107/s2052252519014003.
Shelby, M. L., Gilbile, D., Grant, T. D., Seuring, C., Segelke, B. W., He, W., Evans, A. C., Pakendorf, T., Fischer, P., Hunter, M. S., Batyuk, A., Barthelmess, M., Meents, A., Coleman, M. A., Kuhl, T. L., & Frank, M. A fixed-target platform for serial femtosecond crystallography in a hydrated environment. United States. doi:10.1107/s2052252519014003.
Shelby, M. L., Gilbile, D., Grant, T. D., Seuring, C., Segelke, B. W., He, W., Evans, A. C., Pakendorf, T., Fischer, P., Hunter, M. S., Batyuk, A., Barthelmess, M., Meents, A., Coleman, M. A., Kuhl, T. L., and Frank, M. Wed . "A fixed-target platform for serial femtosecond crystallography in a hydrated environment". United States. doi:10.1107/s2052252519014003.
@article{osti_1605168,
title = {A fixed-target platform for serial femtosecond crystallography in a hydrated environment},
author = {Shelby, M. L. and Gilbile, D. and Grant, T. D. and Seuring, C. and Segelke, B. W. and He, W. and Evans, A. C. and Pakendorf, T. and Fischer, P. and Hunter, M. S. and Batyuk, A. and Barthelmess, M. and Meents, A. and Coleman, M. A. and Kuhl, T. L. and Frank, M.},
abstractNote = {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.},
doi = {10.1107/s2052252519014003},
journal = {IUCrJ},
number = [1],
volume = [7],
place = {United States},
year = {2020},
month = {1}
}

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