Status of the crystallography beamlines at Elettra 2 Andrea Lausi - PowerPoint PPT Presentation

A. Lausi Status of the crystallography beamlines at Elettra

• 2 Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

Photon sources at Elettra 1000 100 10 1 0.1 33 2 /0.1%bw) 10 Wavelength(nm) 31 10 FERMI 10 nm 40 nm 29 10 2 /mrad 100 nm 27 10 Peak Brilliance (Photons/s/mm 25 10 EUFELE 23 10 Elettra 21 U5.6 10 E=2 GeV I=400 mA 1 U12.5 3 19 10 5 FEU Short U5.6 SCW 1 W14.0 3 17 5 10 Short W15.0 Bending Magnet 15 10 0 1 2 3 4 5 10 10 10 10 10 10 Photon Energy (eV) Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

Diffraction at ELETTRA 1995-2006: XRD beamline macromolecules but also material science, high pressure, fibers, inorganic crystals... Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015 4

HRD(1) diffraction beamline (1995 version) refocussing mirror monochromator filter sample stage source slits Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015 5

XRD1: present and future General purpose X-ray beamline with a extended spectrum(4-21 keV) from a wiggler source, shared with the SAXS BL. The BL hosts PX and material science experiments, in collaboration with CNR. Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

XRD1: present and future • Top-up mode introduced serious thermal load problem affecting the BL monochromator. Three water direct-cooled crystals already substituted in the last three years. This was reflected in a reduced (total) flux and enlarged spot size, causing problems both with users and industry (long exposure time, and beam divergence in particular). A closed loop cryogenic system (liquid nitrogen) from Bruker has been purchased and is expected to be tested by the early 2015. XRD1 is expected to be up and running at full capacity at the beginning of 2015. Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

Structural biology lab Molecular biology Large scale protein expression Protein purification Protein characterisation Biophysical analysis Biochemical analysis Crystallisation Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

The PROTEO protein factory Recombinant proteins are fundamental “ reagents ” for basic and applied bioresearch. Obtaining pure and folded proteins is usually a bottleneck in biological studies. X-Domain Y-Domain P R C SCALE UP Multiple Cloning Test Expression Purification Construct Design Thanks to an INTERREG grant we have established a biotechnological platform that provides a specialized support to express and purify recombinant proteins for functional and structural studies . We implemented a multistep process to go from gene to protein with high efficiency using multiparallel protocols in bacteria , insect and mammalian cell expression systems Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

Expression & purification facilities Tecan liquid handler robotic platform for High Troughput cloning & small scale purification 2 Sartorius Certomat BS-1 orbital shakers to grow bacterial cells 5 Sartorius fermentors for controlled expression in prokaryotic and eukaryotic cells , including 2 Biostat B (2L), 2 Biostat C (10L) and one Cultibag 20/50 RM (50 L). 4 Akta FPLC with column fleet for chromatographic protein purification Suite of centrifuges (Beckmann Allegra 22R, Allegra 64R, Avanti J-26XP) with rotor fleet Spectrophotometers & gel imaging system (ImageQuant) Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

Molecular biology lab Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

Bacterial protein expression Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

The crystallization suite Liquid handler Tecan EVO Freedom 150 for re-formatting commercial crystallization screens and setting up crystallisation plates TTP Labtech Mosquito nanoliter crystallization robot to set up 50-100nL crystallization drops Explora Nova Xtal Focus system for automatic crystal imaging and remote monitoring of crystallization experiments The facility has been designed to be open for external users . Users will be able to bring/send purified protein samples, set up a variety of commercial crystallization screens, and monitor the results of the experiments in remote, through a web- based system. Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

Crystallization lab Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

2009 - Present: MCX new XRD1 BM 7.1 currently in construction: XRD2 cluster Beamline designed for non  single crystal diffraction experiments Open to users since 2009  Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

Before MCX: “ Powder Diffraction Facilities at ELETTRA ” • Powder in capillary on XRD1 area detector (Si powder @ ~18 keV) • Fit2D to integrate rings • Resolution limited by spatial response of detector and by sample dimensions Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015 16

Diffraction analyses  Phase identifications  Texture analysis  Phase transitions  Crystal structure determination  Crystal structure refinements  Quantitative phase analysis (and crystallinity determination)  Microstructural analyses (crystallite sizes - microstrain)  Residual stress analysis Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015 17

Design Guidelines • High-flux tuneable source with a wide spectral range, (6-12 keV) • Possibility to extend the • Flexible optics – line focus (10 x 1 mm 2 ) – point focus (1 x 1 mm 2 and below) • Accommodate large volume samples • Use of different detectors • Sample temperature control • User-friendly software Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015 18

MCX optical design : source Photons/s/0.1%bw (/2 mrad horiz /0.27 mrad vert.) ELETTRA operated at 2.4 GeV, 200 mA bending magnet spectral flux Flux at the sample (Air environment) Flux at the sample (He environment) 10 13 10 13 Photons/s 10 12 10 12 10 11 10 11 10 10 10 10 2 3 4 5 6 7 8 9 2 10 3 10 4 Photon Energy (eV) l (Å ) 9 8 7 6 5 4 3 2 9 8 7 6 10 1 Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015 19

MCX optical design : source Photons/s/0.1%bw (/2 mrad horiz /0.27 mrad vert.) ELETTRA operated at 2.4 GeV, 200 mA bending magnet spectral flux Flux at the sample (Air environment) Flux at the sample (He environment) 10 13 10 13 Photons/s 10 12 10 12 10 11 10 11 10 10 10 10 2 3 4 5 6 7 8 9 2 10 3 10 4 Photon Energy (eV) l (Å ) 9 8 7 6 5 4 3 2 9 8 7 6 10 1 Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015 20

MCX optical design : outline • High-transmission Filter- window assembly • Collimating Pre-mirror • Wide range monochromator with II crystal bender for dynamical sagittal focusing • Bendable focusing mirror Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015 21

Sagittal Focussing Grazing incidence reflection optics: • geometrical loss • small acceptances • difficult to manufacture • expensive bent II crystal Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015 22

The Elettra Hard X-ray Monochromator long 2nd crystal movement capability allow for the wide E. Busetto, I. Cudin, G. Fava R. Borghes, G. Cautero energy range Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015 23

The Elettra Hard X-ray Monochromator Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015 24

Front-end Hutch The CX project … Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin Anrea Lausi 20-21 April 2015 25

First mirror • 26 Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

First mirror 0.5 µ rad slope error measured on the Elettra Long Trace Profilometer • 27 Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

0.65 µ rad slope error • 28 Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

From the Optical to the Experimental Hutch • 29 Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

Experimental Station • 30 Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

Experimental Station 4 circle diffractometer  (0.0001 ° precision in 2 θ) XYZ translation stage  Recieving slits  Analyzer crystal / scintillator  detection system Hot air blower (up to 1273K)  Cryo stream (down to 100K)  Laser sensor for accurate  sample positioning • 31 Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

General User Interface Control system based on pyton General command interface for: • driving motors • theta-2theta scan • multiple theta-2theta scans • single or two motor scan • multiple scans • monochromator functions • calibration functions • warnings management... • ... • 32 Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015

Experimental Station • 33 Andrea Lausi – 7 ° ILSF Users ’ Meeting – Qazvin 20-21 April 2015