Fixed-target protein serial microcrystallography with an x-ray free electron laser
We present results from experiments at the Linac Coherent Light Source (LCLS) demonstrating that serial femtosecond crystallography (SFX) can be performed to high resolution (~2.5 Å) using protein microcrystals deposited on an ultra-thin silicon nitride membrane and embedded in a preservation medium...
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| Format: | Online |
| Language: | English |
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Nature Publishing Group
2014
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4129423/ |
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pubmed-4129423 |
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pubmed-41294232014-08-14 Fixed-target protein serial microcrystallography with an x-ray free electron laser Hunter, Mark S. Segelke, Brent Messerschmidt, Marc Williams, Garth J. Zatsepin, Nadia A. Barty, Anton Benner, W. Henry Carlson, David B. Coleman, Matthew Graf, Alexander Hau-Riege, Stefan P. Pardini, Tommaso Seibert, M. Marvin Evans, James Boutet, Sébastien Frank, Matthias Article We present results from experiments at the Linac Coherent Light Source (LCLS) demonstrating that serial femtosecond crystallography (SFX) can be performed to high resolution (~2.5 Å) using protein microcrystals deposited on an ultra-thin silicon nitride membrane and embedded in a preservation medium at room temperature. Data can be acquired at a high acquisition rate using x-ray free electron laser sources to overcome radiation damage, while sample consumption is dramatically reduced compared to flowing jet methods. We achieved a peak data acquisition rate of 10 Hz with a hit rate of ~38%, indicating that a complete data set could be acquired in about one 12-hour LCLS shift using the setup described here, or in even less time using hardware optimized for fixed target SFX. This demonstration opens the door to ultra low sample consumption SFX using the technique of diffraction-before-destruction on proteins that exist in only small quantities and/or do not produce the copious quantities of microcrystals required for flowing jet methods. Nature Publishing Group 2014-08-12 /pmc/articles/PMC4129423/ /pubmed/25113598 http://dx.doi.org/10.1038/srep06026 Text en Copyright © 2014, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by-nc-nd/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/ |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Hunter, Mark S. Segelke, Brent Messerschmidt, Marc Williams, Garth J. Zatsepin, Nadia A. Barty, Anton Benner, W. Henry Carlson, David B. Coleman, Matthew Graf, Alexander Hau-Riege, Stefan P. Pardini, Tommaso Seibert, M. Marvin Evans, James Boutet, Sébastien Frank, Matthias |
| spellingShingle |
Hunter, Mark S. Segelke, Brent Messerschmidt, Marc Williams, Garth J. Zatsepin, Nadia A. Barty, Anton Benner, W. Henry Carlson, David B. Coleman, Matthew Graf, Alexander Hau-Riege, Stefan P. Pardini, Tommaso Seibert, M. Marvin Evans, James Boutet, Sébastien Frank, Matthias Fixed-target protein serial microcrystallography with an x-ray free electron laser |
| author_facet |
Hunter, Mark S. Segelke, Brent Messerschmidt, Marc Williams, Garth J. Zatsepin, Nadia A. Barty, Anton Benner, W. Henry Carlson, David B. Coleman, Matthew Graf, Alexander Hau-Riege, Stefan P. Pardini, Tommaso Seibert, M. Marvin Evans, James Boutet, Sébastien Frank, Matthias |
| author_sort |
Hunter, Mark S. |
| title |
Fixed-target protein serial microcrystallography with an x-ray free electron laser |
| title_short |
Fixed-target protein serial microcrystallography with an x-ray free electron laser |
| title_full |
Fixed-target protein serial microcrystallography with an x-ray free electron laser |
| title_fullStr |
Fixed-target protein serial microcrystallography with an x-ray free electron laser |
| title_full_unstemmed |
Fixed-target protein serial microcrystallography with an x-ray free electron laser |
| title_sort |
fixed-target protein serial microcrystallography with an x-ray free electron laser |
| description |
We present results from experiments at the Linac Coherent Light Source (LCLS) demonstrating that serial femtosecond crystallography (SFX) can be performed to high resolution (~2.5 Å) using protein microcrystals deposited on an ultra-thin silicon nitride membrane and embedded in a preservation medium at room temperature. Data can be acquired at a high acquisition rate using x-ray free electron laser sources to overcome radiation damage, while sample consumption is dramatically reduced compared to flowing jet methods. We achieved a peak data acquisition rate of 10 Hz with a hit rate of ~38%, indicating that a complete data set could be acquired in about one 12-hour LCLS shift using the setup described here, or in even less time using hardware optimized for fixed target SFX. This demonstration opens the door to ultra low sample consumption SFX using the technique of diffraction-before-destruction on proteins that exist in only small quantities and/or do not produce the copious quantities of microcrystals required for flowing jet methods. |
| publisher |
Nature Publishing Group |
| publishDate |
2014 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4129423/ |
| _version_ |
1613123253816000512 |