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Method Article
Alke Meents
Deutsches Elektronen Synchrotron DESY, Photon Science, Notkestrasse 85, D-22607 Hamburg, Germany; Center for Free Electron Laser Science (CFEL), Luruper Chaussee 149, D-22761 Hamburg, Germany.
Corresponding Author
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We have developed a protocol for high-speed fixed-target serial femtosecond crystallography (SFX) at X-ray free electron lasers (XFELs). Using the Roadrunner goniometer for fast scanning of microcrystals loaded on a micro-patterned silicon chip, our method allows for usage of the full 120 Hz repetition rate of the Linear Coherent Light Source (LCLS). By synchronizing the sample exchange to the arrival of the LCLS X-ray pulses our approach results in most efficient use of sample material and beamtime. The presented protocol describes the loading of microcrystals on micro-patterned silicon chips and subsequent data collection using the Roadrunner goniometer installed at an XFEL beamline. Data collection can be performed either at cryogenic or room temperature. We further describe the Roadrunner control and data acquisition software and data processing using cppxfel. Due to the small sample amounts required and the low background scattering levels achievable, the method is ideally suited for data collection of precious microcrystals from viruses or membrane proteins, where often only limited amounts are available. This protocol accompanies Roedig P et al, Nature Methods (10.1038/nmeth.4335, published online June 19, 2017).
Keywords
fixed-target serial femtosecond crystallography, protein crystallography, virus crystallography
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A.M. is a CEO and shareholder of DESY spin-off company Suna-Precision GmbH. T.P. and B.R. are also shareholders. Suna-Precision GmbH sells technical equipment for experiments with X-rays including different micro-structured silicon chips for serial crystallography experiments. I.V. is shareholder and CTO of the company FinnLitho, which produced the silicon chips used in the experiment and described in the protocol.
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.txt
List 1 Exemplary parameter input file as used for indexing with cppxfel Shown parameters were used for indexing of BEV2 XFEL data as described in the associated publication to this protocol.
- supplement0.txt
List 3 Exemplary parameter input file as used for post-refinement with cppxfel Shown parameters were used for post-refinement of BEV2 XFEL data as described in the associated publication to this protocol.
- supplement0.txt
List 2 Exemplary parameter input file as used for initial orientation matrix refinement with cppxfel Shown parameters were used for initial orientation matrix refinement of BEV2 XFEL data as described in the associated publication to this protocol.
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Associated Publications
High-speed fixed-target serial virus crystallography
by Philip Roedig, Helen M Ginn, Tim Pakendorf, +14
Nature Methods (28 June, 2017)
Method Article
Alke Meents
Deutsches Elektronen Synchrotron DESY, Photon Science, Notkestrasse 85, D-22607 Hamburg, Germany; Center for Free Electron Laser Science (CFEL), Luruper Chaussee 149, D-22761 Hamburg, Germany.
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 29 Jun, 2017
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
We have developed a protocol for high-speed fixed-target serial femtosecond crystallography (SFX) at X-ray free electron lasers (XFELs). Using the Roadrunner goniometer for fast scanning of microcrystals loaded on a micro-patterned silicon chip, our method allows for usage of the full 120 Hz repetition rate of the Linear Coherent Light Source (LCLS). By synchronizing the sample exchange to the arrival of the LCLS X-ray pulses our approach results in most efficient use of sample material and beamtime. The presented protocol describes the loading of microcrystals on micro-patterned silicon chips and subsequent data collection using the Roadrunner goniometer installed at an XFEL beamline. Data collection can be performed either at cryogenic or room temperature. We further describe the Roadrunner control and data acquisition software and data processing using cppxfel. Due to the small sample amounts required and the low background scattering levels achievable, the method is ideally suited for data collection of precious microcrystals from viruses or membrane proteins, where often only limited amounts are available. This protocol accompanies Roedig P et al, Nature Methods (10.1038/nmeth.4335, published online June 19, 2017).
Figure 1
Figure 2
Figure 3
Figure 4
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Associated Publications
High-speed fixed-target serial virus crystallography
by Philip Roedig, Helen M Ginn, Tim Pakendorf, +14
Nature Methods (28 June, 2017)