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  Viscous hydrophilic injection matrices for serial crystallography

Kovácsová, G., Grünbein, M. L., Kloos, M., Barends, T., Schlesinger, R., Heberle, J., et al. (2017). Viscous hydrophilic injection matrices for serial crystallography. IUCrL Internationale Union für Kristallographie, 4(4), 400-410. doi:10.1107/S2052252517005140.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002E-294E-F Version Permalink: http://hdl.handle.net/21.11116/0000-0000-2A29-4
Genre: Journal Article

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IUCrJ_4_2017_400.pdf (Any fulltext), 491KB
 
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 Creators:
Kovácsová, Gabriela1, Author              
Grünbein, Marie Luise1, Author              
Kloos, Marco1, Author              
Barends, Thomas1, Author              
Schlesinger, Ramona, Author
Heberle, Joachim, Author
Kabsch, Wolfgang1, Author              
Shoeman, Robert L.1, Author              
Doak, Bruce1, Author              
Schlichting, Ilme1, Author              
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1Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Max Planck Society, ou_1497700              

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Free keywords: high-throughput serial crystallography; room-temperature crystallography; microcrystal injection; XFEL; high-viscosity extrusion.
 Abstract: Serial (femtosecond) crystallography at synchrotron and X-ray free-electron laser (XFEL) sources distributes the absorbed radiation dose over all crystals used for data collection and therefore allows measurement of radiation damage prone systems, including the use of microcrystals for room-temperature measurements. Serial crystallography relies on fast and efficient exchange of crystals upon X-ray exposure, which can be achieved using a variety of methods, including various injection techniques. The latter vary significantly in their flow rates - gas dynamic virtual nozzle based injectors provide very thin fast-flowing jets, whereas high-viscosity extrusion injectors produce much thicker streams with flow rates two to three orders of magnitude lower. High-viscosity extrusion results in much lower sample consumption, as its sample delivery speed is commensurate both with typical XFEL repetition rates and with data acquisition rates at synchrotron sources. An obvious viscous injection medium is lipidic cubic phase (LCP) as it is used for in meso membrane protein crystallization. However, LCP has limited compatibility with many crystallization conditions. While a few other viscous media have been described in the literature, there is an ongoing need to identify additional injection media for crystal embedding. Critical attributes are reliable injection properties and a broad chemical compatibility to accommodate samples as heterogeneous and sensitive as protein crystals. Here, the use of two novel hydro-gels as viscous injection matrices is described, namely sodium carb-oxy-methyl cellulose and the thermo-reversible block polymer Pluronic F-127. Both are compatible with various crystallization conditions and yield acceptable X-ray background. The stability and velocity of the extruded stream were also analysed and the dependence of the stream velocity on the flow rate was measured. In contrast with previously characterized injection media, both new matrices afford very stable adjustable streams suitable for time-resolved measurements.

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Language(s): eng - English
 Dates: 2017-03-092017-04-112017-05-052017-07
 Publication Status: Published in print
 Pages: 11
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 Table of Contents: -
 Rev. Method: Peer
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Title: IUCrL Internationale Union für Kristallographie
Source Genre: Journal
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Publ. Info: England : Chester
Pages: - Volume / Issue: 4 (4) Sequence Number: - Start / End Page: 400 - 410 Identifier: ISSN: 2052-2525