title: Versatile sample environments and automation for biological solution X-ray scattering experiments at the P12 beamline (PETRA III, DESY).
abstract: A high-brilliance synchrotron P12 beamline of the EMBL located at the PETRA III storage ring (DESY, Hamburg) is dedicated to biological small-angle X-ray scattering (SAXS) and has been designed and optimized for scattering experiments on macromolecular solutions. Scatterless slits reduce the parasitic scattering, a custom-designed miniature active beamstop ensures accurate data normalization and the photon-counting PILATUS 2M detector enables the background-free detection of weak scattering signals. The high flux and small beam size allow for rapid experiments with exposure time down to 30-50 ms covering the resolution range from about 300 to 0.5 nm. P12 possesses a versatile and flexible sample environment system that caters for the diverse experimental needs required to study macromolecular solutions. These include an in-vacuum capillary mode for standard batch sample analyses with robotic sample delivery and for continuous-flow in-line sample purification and characterization, as well as an in-air capillary time-resolved stopped-flow setup. A novel microfluidic centrifugal mixing device (SAXS disc) is developed for a high-throughput screening mode using sub-microlitre sample volumes. Automation is a key feature of P12; it is controlled by a beamline meta server, which coordinates and schedules experiments from either standard or nonstandard operational setups. The integrated SASFLOW pipeline automatically checks for consistency, and processes and analyses the data, providing near real-time assessments of overall parameters and the generation of low-resolution models within minutes of data collection. These advances, combined with a remote access option, allow for rapid high-throughput analysis, as well as time-resolved and screening experiments for novice and expert biological SAXS users.
authors.affiliation: European Molecular Biology Laboratory, Hamburg Outstation , Notkestrasse 85, Hamburg, 22603, Germany.
authors.forename: Clement E
authors.initials: CE
authors.lastname: Blanchet
authors.affiliation: European Molecular Biology Laboratory, Hamburg Outstation , Notkestrasse 85, Hamburg, 22603, Germany.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Spilotros
authors.affiliation: Laboratory for MEMS Applications, IMTEK - Department of Microsystems Engineering, University of Freiburg , Georges-Koegler-Allee 103, Freiburg, 79110, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Schwemmer
authors.affiliation: European Molecular Biology Laboratory, Hamburg Outstation , Notkestrasse 85, Hamburg, 22603, Germany.
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Graewert
authors.affiliation: European Molecular Biology Laboratory, Hamburg Outstation , Notkestrasse 85, Hamburg, 22603, Germany.
authors.forename: Alexey
authors.initials: A
authors.lastname: Kikhney
authors.affiliation: European Molecular Biology Laboratory, Hamburg Outstation , Notkestrasse 85, Hamburg, 22603, Germany.
authors.forename: Cy M
authors.initials: CM
authors.lastname: Jeffries
authors.affiliation: European Molecular Biology Laboratory, Hamburg Outstation , Notkestrasse 85, Hamburg, 22603, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Franke
authors.affiliation: Laboratory for MEMS Applications, IMTEK - Department of Microsystems Engineering, University of Freiburg , Georges-Koegler-Allee 103, Freiburg, 79110, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Mark
authors.affiliation: Laboratory for MEMS Applications, IMTEK - Department of Microsystems Engineering, University of Freiburg , Georges-Koegler-Allee 103, Freiburg, 79110, Germany.
authors.forename: Roland
authors.initials: R
authors.lastname: Zengerle
authors.affiliation: European Molecular Biology Laboratory, Grenoble Outstation , 6 rue Jules Horowitz, BP 181, Grenoble, 38042, France.
authors.forename: Florent
authors.initials: F
authors.lastname: Cipriani
authors.affiliation: European Molecular Biology Laboratory, Hamburg Outstation , Notkestrasse 85, Hamburg, 22603, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Fiedler
authors.affiliation: European Molecular Biology Laboratory, Hamburg Outstation , Notkestrasse 85, Hamburg, 22603, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Roessle
authors.affiliation: European Molecular Biology Laboratory, Hamburg Outstation , Notkestrasse 85, Hamburg, 22603, Germany.
authors.forename: Dmitri I
authors.initials: DI
authors.lastname: Svergun
keywords: automated hardware and software systems; biological small-angle X-ray scattering; high-brilliance P12 synchrotron beamline
references.citation: J Synchrotron Radiat. 2015 Mar;22(2):461-4
references.pmid: 25723949
references.citation: Cell Mol Biol (Noisy-le-grand). 2000 Jul;46(5):993-1004
references.pmid: 10976879
references.citation: Protein Sci. 2002 Jan;11(1):72-81
references.pmid: 11742124
references.citation: J Appl Crystallogr. 2012 Apr 1;45(Pt 2):213-223
references.pmid: 22509071
references.citation: Biophys J. 1999 Jun;76(6):2879-86
references.pmid: 10354416
references.citation: Chem Rev. 2001 Jun;101(6):1727-61
references.pmid: 11709997
references.citation: Acta Crystallogr D Biol Crystallogr. 2013 Oct;69(Pt 10):2072-80
references.pmid: 24100325
references.citation: Annu Rev Biophys Biomol Struct. 2007;36:307-27
references.pmid: 17284163
references.citation: Biomicrofluidics. 2008 Jun 06;2(2):24104
references.pmid: 19693407
references.citation: J Colloid Interface Sci. 2007 Feb 15;306(2):379-85
references.pmid: 17141795
references.citation: Biochemistry. 1990 Mar 13;29(10):2564-72
references.pmid: 2110472
references.citation: Chem Rev. 2006 Aug;106(8):3031-42
references.pmid: 16895316
references.citation: Proc Natl Acad Sci U S A. 2013 Apr 23;110(17):6706-11
references.pmid: 23569240
references.citation: Biopolymers. 2011 Aug;95(8):550-8
references.pmid: 21442608
references.citation: Curr Opin Struct Biol. 2013 Oct;23(5):748-54
references.pmid: 23835228
references.citation: J Synchrotron Radiat. 2015 Mar;22(2):273-9
references.pmid: 25723929
references.citation: J Appl Crystallogr. 2008 Aug 16;41(Pt 5):913-917
references.pmid: 25484841
references.citation: J Synchrotron Radiat. 2004 Jul 1;11(Pt 4):314-8
references.pmid: 15211037
references.citation: Biochemistry. 1992 Sep 8;31(35):8329-35
references.pmid: 1525171
references.citation: J Synchrotron Radiat. 2013 Nov;20(Pt 6):820-5
references.pmid: 24121320
references.citation: J Synchrotron Radiat. 2013 Jul;20(Pt 4):660-4
references.pmid: 23765312
references.citation: Science. 2011 Oct 28;334(6055):517-20
references.pmid: 22034434
references.citation: Phys Med Biol. 2002 Feb 21;47(4):577-92
references.pmid: 11900192
references.citation: J Synchrotron Radiat. 2003 May 1;10(Pt 3):287-8
references.pmid: 12714764
references.citation: Nucleic Acids Res. 2014 May;42(9):5978-92
references.pmid: 24634441
references.citation: Acta Crystallogr D Biol Crystallogr. 2015 Jan 1;71(Pt 1):67-75
references.pmid: 25615861
references.citation: PLoS Biol. 2007 May;5(5):e134
references.pmid: 17472440
references.citation: J Synchrotron Radiat. 2010 Nov;17(6):774-81
references.pmid: 20975223
references.citation: J Mol Biol. 1991 Oct 5;221(3):941-59
references.pmid: 1942038
references.citation: Annu Rev Phys Chem. 2013;64:37-54
references.pmid: 23216378
references.citation: J Appl Crystallogr. 2009 Apr 1;42(Pt 2):342-346
references.pmid: 27630371