Robotic sample changer at P12

Robotic sample changer at P12

The call for the 2020 beamtime proposals is now closed. Possibilities for rolling proposals submission, if/when appear, will be announced.

EMBL Hamburg operates a synchrotron beamline P12 for small angle X-ray scattering (SAXS) at the PETRA III storage ring, under the coordination of Dmitri Svergun.

P12 is an undulator beamline utilizing the wavelength range between 0.6 and 3 Å. The beamline is optimised for low background data collection from macromolecular solutions and kinetic experiments with high temporal resolution. The endstation is equipped with photon counting Pilatus and Eiger detectors, a robotic sample changer and in-line size-exclusion chromatography (SEC-SAXS) setups. P12 allows for fully automated and remote data acquisition and processing using the Beamline Meta Server software. Mail-in access is encouraged, both for sample changer and SEC-SAXS projects.

Access to the beamline P12 is available to all research groups, and is prioritised on scientific grounds only. Project proposals for beamline access are evaluated by the Project Evaluation Committee (PEC). For guidance on writing a project proposal for SAXS beam time, please consult these instructions.

Further enquiries may be made to saxs@embl-hamburg.de.

How to reference P12

The use of the EMBL beamlines must be acknowledged in all publications or presentations arising from data collection at the EMBL Hamburg unit. The following note is expected to appear in the acknowledgements, giving reference to the station(s) used, supporting beamline staff or any other assistance or information, as appropriate, e.g. EU contract numbers, proposal numbers.

“The synchrotron SAXS data was collected at beamline P12 operated by EMBL Hamburg at the PETRA III storage ring (DESY, Hamburg, Germany). We would like to thank N.N.** for the assistance in using the beamline.”


**If appropriate, names of the beamline staff in case they are not co-authors.

In case of P12, please cite the beamline paper:

Blanchet CE, Spilotros A, Schwemmer F, Graewert MA, Kikhney A, Jeffries CM, Franke D, Mark D, Zengerle R, Cipriani F, Fiedler S, Roessle M, Svergun DI. (2015) J Appl Crystallogr. 48(2): 431-443. [Link to Open Access]
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4379436/

Experimental possibilities
  • SAXS experiments on macromolecules, nanoparticles and bio-nanoconjugates ranging from a few kDa to Gda.
  • High throughput data collection using an automated liquid handling sample changer using about 30 μl sample per measurement (in-vacuum setup).
  • A single frame exposure time is 45 msec (with a standard monochromator) or 1.2 msec (multilayer monochromator) leading to total exposures about 1-2 seconds depending on the capillary diameter (1 or 2 mm), flow rate, and susceptibility to radiation damage.
  • Size-exclusion (SEC-SAXS), ion-exchange and affinity chromatography SAXS (FPLC/HPLC – Agilent BioInert system with an autosampler).
  • Parallel SEC-SAXS measurements combined with on-line multi-angle laser light scattering (MALLS) and dynamic light scattering (Wyatt TREOS-QELS/T-Rex).
  • Wide-angle scattering (WAXS) setup up to s≈25nm-1 (nominal resolution 0.25nm)
  • Time resolved measurements (sub-msec resolution with multilayer monochromator) and temperature scans.
  • A stopped flow device (BioLogic, in-air setup).
  • Sample holders for solutions, gels, films, powders (in-air set up).
  • ‘Bring-your-own’ user-supplied in-air sample environments.
  • Mail-in operations with the possibility of remote access mode.
  • Additional off-line dynamic light scattering (Wyatt NanoStar) and Refractive index (Anton Paar Abbemat 550) instruments.
Parameters
Synchrotron source Standard Petra U29 undulator
Monochromator

Oxford double crystal monochromator, crystal Si (111) or Double Multilayer monochromator (DMM)
Mirror KB bimorph adaptative mirrors, Rh (-coated) and SiO3 (uncoated)
Control system Beckhoff Twincat, TINE
Wavelength 0.06 – 0.3 nm (DCM), 0.08 -0.17 (DMM)
Focal spot 0.2 mm x 0.12 mm
Flux 1013 ph/s (DCM), 5*1014 (DMM)
Slits 4*2 pairs: two sets of Cinel water cooled slits, defining slits: motorized highly polished tungsten blade, guard slits: Hybrid Scatterless slits (Xenocs)
Automated sample changer 2 x 96 wells (crystallization plates)
4 x 8 samples (200 µl PCR stripes) + 12 buffers (1.5 ml Eppendorf tubes)
temperature controlled storage (5-40°C)
SEC-SAXS

FPLC/HPLC – Agilent BioInert system with autosampler
On-line multi-angle laser light scattering (MALLS) and dynamic light scattering (Wyatt TREOS-QELS/T-Rex).
Exposure unit Quartz capillary, diameter 1 or 2 mm, automated filling and rinsing, temperature controlled (7-50°C). Filling volume: 30-40 µl for standard continuous flow operations (down to 5 µl for static measurements).
Alternative sample environments YZ- alignment stage for the positioning of user-made sample cell/holder.
Temperature controlled (0-100°C) capillary holder (Ø 2mm).
Stopped flow SFM-400 (Bio-Logic)
Detectors Photon counting Pilatus 6M (423.6 x 434.6mm2), Pilatus 2M pixel detector (253 x 288 mm2), Pilatus 1M-W detector (423.6 x 70 mm2), Eiger 4M detector (155.2 x 162.5 mm2, 750 Hz frame rate)
Sample-detector distance Tunable: 1.6 to 6.1 m