Status and developments in BL13-XALOC at Alba

Why XALOC Difractòmetre Orienta la mostra Robot Posiciona la mostra Detector Flux congelador a -173º C Microscopi XALOC-BL13 beamline determines the 3D structure of macromolecules (proteins, ADN/ARN) using X-ray crystallography 0,1 mm Detector Cabina Experimental de Xaloc-BL13

3D image at atomic resolution of a protein+DNA 3. Solve a X-ray diffraction pattern of protein crystal at the pixel detector Protein crystal 3D image at atomic resolution of a protein+DNA

High demand User Operation Cycle 1 (2012+) Cycle 2 (2013) 97 BT days 34 Udays (29 proposals) Users can not manage LN2, safety training is not yet on-line, improve User office portal (introduce coworkers), user accounts ~50 proposals

Beamline status Beamline status : Operative (but not completed yet) Very positive comments from all (!) users Typical beam size at sample is 5010 um FWHM (HV) as seen on a 20-um fluorescence screen Unfocused and defocused modes were tested although not currently used. Very reliable beam optics (alignment done daily in 10’) Beam at sample drifts (typically downwards 1 um/hour) Beam recovered easily after a shutdown. Beam aligned using EH positioning tables. Optics (mirror pitches) could also be used. Measured beam flux is >2·1012 ph/s/250mA 5010 mm (FWHM, HV) Flux (ph/s/250mA) E (keV) #3 #5 #7 #9 #11 #13 Underlying: diagnostics are very important

Vertical Focusing Mirror Horizontal Focusing Mirror Beamline description Beamline concept : In-vacuum Undulator IVU21.6 + Channel-cut Si(111) monochromator + KB focusing mirrors Beamline status : Operative Vertical Focusing Mirror Mono- chromator Horizontal Focusing Mirror Diffractometer Detector Distance from source (m) 20.100 Sample changer 22.800 24.785 Dv = 3.47 29.375 Dh = 5.4 Experimental Hutch

Horizontal Focusing Mirror Vertical Focusing Mirror XALOC Layout 29.375 m Detector Diffractometer 24.785 m Dh = 5.4 Photon shutter Horizontal Focusing Mirror 22.800 m Dv = 3.47 Vertical Focusing Mirror 20.100 m Sample changer Monochromator Attenuator Other optical elements Fluorescence screens Diode XBPMs Slits FE trigger unit

Optics Channel cut Si(111), 6 mm gap, by CINEL (crystals by Crystal Scientific) 5-22 keV, with 1 mm excursion (Pt-K →Se-K: 30 mm; complete Se-MAD: 11 mm) 2nd xtal pitch (stepper motor) 2nd xtal pitch V feedback (piezo) Bremsstrahlung block I0 diode beam channel cut crystal cryogenic pipes accelerometer Resolution Bragg Repeatability Bragg 1.10 ± 0.08 mrad (0.064 mdeg) 2 mrad monodirectional 4 mrad bidirectional Measured at 1 full step In range 5-23 deg Measures limited by thermal drift

Horizontal Focusing Mirror Optics: mirrors 2 mirrors, in a Kirkpatrick-Baez configuration Manufactured by IRELEC, surfaces made by InSync Elliptically bent using 2 motors, equipped with strain gauges Equipped with accelerometers to monitor pitch vibrations Fully installed VFM HFM Optical length 300 mm 600 mm Stripes Si, Rh, Ir Z Roll Pitch/Z Range 10 mm 15 mrad Tbd Resolution 0.08 mm 1 mrad - Repeatability 4 mm 1 mm tbd Vertical Focusing Mirror Horizontal Focusing Mirror

Lysozyme data set at Rsym (inner shell)<5% High quality data: Lysozyme Test lysozyme crystal at 100K. Excellent overall statistics: Rsym = 3.8 % (90 deg) Also at the lowest resolution shell (2.76A): Rsym= 2 %. Resolution 1.26 Å. Molecular replacement rendered excellent ED maps. lysozyme (a=b=79 Å c=37 Å; P43212) sample-det distance=200 mm l=0.9795 Å, exp time =1 s, Dw = 1° Atomic resolution data: T98D-SH3 domain of c-Src Tyr kinase P3121 , a = b =31.5 Å, c = 106.8 Å First data set collected at 12.6 keV and at 0.93 Å resolution: 0.5 deg oscillation, 90 deg, 1.3 sec exp time Rsym = 3.8 % (90 deg) Second data set collected at 15 keV and at 0.96 Å resolution: 0.5 deg oscillation, 205 deg, 2.6 sec exp time Rsym= 3.9 % P3121 31.5 31.5 106.8 Crystals from A. Cámara, UA, Almería

Solve a structure using MAD at Se-K edge SAD data collection: meganuclease I-Dmol D. Mobilis First ab initio solved structure, by Se K-edge SAD Protein-DNA complex (PDB 2VS8) Se-Met SAD 360˚, 2.4 Å resolution, 1 sec exp time P21, a = 106.6, b = 70.3, c = 107.1, b = 119.9 Protein DNA Fluorescence scan widget Crystals from G. Montoya, CNIO, Madrid

Solve a structure with a unit cell par. a>400 Å Human Rhinovirus 2 Monoclinic cell a = 470 Å, b= 376 Å, c = 465 Å = 90˚, = 99˚, = 90˚ Diffraction to 4 Å. Crystal-detector distance 750 cm (max. distance 1350 cm) Crystals from N. Verdaguer, IBMB, Barcelona

Detectors at the Experimental Hutch Fluorescence Si-drift detector Diamond X-ray position monitors Pixel main detector PIN Diodes and YAG fluorescence screens

Detector Pilatus 6M (Dectris) Photon counting pixel detector Preliminary tests on integration into BL successfully passed Currently ending manufacturing phase Estimated delivery in May 2011 Data transfer and storage system to be defined ALBA turn = 893 ns 124 ns Area 431 x 448 mm2 Pixel size 172 mm Point spread 1 pixel Max. framing rate 12 Hz Dynamic Range 20 bits (106 ct) Usable count rate 2x106 cts/s Energy trimming Yes, recommended E0/2 Gain 3, Fast, standard, high

? ? Detector. What else? 320 um Silicon: needed thicker Smaller pixel size Tiling is tolerable, but gaps should be reduced 2.3 ms dead time: can be smaller? Area, area Triggers, triggers, triggers ALBA turn = 893 ns 124 ns ? Area 431 x 448 mm2 Pixel size 172 mm Point spread 1 pixel Max. framing rate 12 Hz Dynamic Range 20 bits (106 ct) Usable count rate 2x106 cts/s Energy trimming Yes, recommended E0/2 Gain 3, Fast, standard, high ?

Other projects Transmissive PIN diode Collaboration with CNM Pinhole 10x10mm2 transmissive (10 um thick) Pinhole Heavy Z material, 80 um thick Hole 3-20 um diameter High precision tools Sharp tips <1um Slit blades Fluorescence dot Mounted in a sample holder 1 um diameter Used to position beam and spindle

The XALOC Team Engineering Computing Experiments Carles Colldelram Mech Engineer Alex Enrique Mech Technician Fabien Rey Alignment Marcos Quispe Cryogenics Computing Guifré Cuní Controls Engineer Oscar Matilla Electronic Engineer Julio Lidón Xavi Fariña Elec technician Experiments Jordi Juanhuix Jordi Benach Fernando Gil J.C. Martínez metrology Josep Nicolás mirrors, metrology Igors Sics diagnostics First data set April 2012 First Official Users 18 July 2012 Thank you for your attention