Download ppt - IFAE-Alba meeting | Alba, 13 december, 2012XALOC-BL13 1 of 16 Status and developments in BL13-XALOC at Alba

IFAE-Alba meeting | Alba, 13 december, 2012XALOC-BL13 1 of 16

Status and developments in

BL13-XALOC at Alba


Why XALOC

XALOC-BL13 beamline determines the 3D structure of macromolecules (proteins, ADN/ARN) using X-ray crystallography

0,1 mm

Detector

DifractòmetreOrienta la mostra

RobotPosiciona la mostra

MicroscopiFlux congelador a -173º C


3. Solve a

Protein crystal

X-ray diffraction patternof protein crystal at the pixel detector

3D image at atomic resolution of a protein+DNA


High demand

January February March April May June July August September October November DecemberShift Shift Shift Shift Shift Shift Shift Shift Shift Shift Shift Shift

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34 Udays (29 proposals)

53 BT days

Cycle 1 (2012+) Cycle 2 (2013)

~50 proposals

• User Operation

97 BT days


Beamline status

• Typical beam size at sample is 5010 um FWHM (HV) 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

Very positive comments from all (!) users

5010 m(FWHM, HV)

Beamline status : Operative (but not completed yet)

Flux

(ph

/s/2

50m

A)

E (keV)

#3#5

#7

#9

#11

#13

Underlying: diagnostics are very important


Mono- chromator

Vertical Focusing Mirror

Dh = 5.4Dv = 3.47

Beamline description

Horizontal Focusing Mirror

Diffractometer

Sample changer

Detector

29.375

24.78522.800

20.100

Distance from source (m)

Beamline concept : In-vacuum Undulator IVU21.6 + Channel-cut Si(111) monochromator + KB focusing mirrors

ExperimentalHutch

Beamline status : Operative


Monochromator



Attenuator

FE trigger unit

Fluorescence screens

Diode XBPMs

Detector

Diffractometer

Sample changer

Slits

Photon shutter

Other optical elements

29.375 m

24.785 m

22.800 m

20.100 m

Dh = 5.4

Dv = 3.47

XALOC Layout


• 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 m; complete Se-MAD: 11 m)

cryogenic pipes

2nd xtal pitch (stepper motor)

2nd xtal pitchV feedback (piezo)

Bremsstrahlung block

channel cut crystal

Resolution Bragg Repeatability Bragg

1.10 ± 0.08 rad(0.064 mdeg)

2 rad monodirectional4 rad bidirectional

Measured at 1 full stepIn range 5-23 deg

Measures limited by thermal drift In range 5-23 deg

I0 diode

beam

accelerometer

Optics


• 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 Si, Rh, Ir

Z Roll Pitch/Z

Range 10 mm 15 mrad 10 mm Tbd

Resolution 0.08 m 1 rad - TbdRepeatability 4 m 1 rad 1 m tbd

Optics: mirrors


Lysozyme data set at Rsym (inner shell)<5%

• 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=0.9795 Å, exp time =1 s, = 1°

High quality data: Lysozyme

• 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 timeRsym = 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 %

Atomic resolution data:T98D-SH3 domain of c-Src Tyr kinase

P3121 31.5 31.5 106.8Crystals from A. Cámara, UA, Almería


Protein

DNA

• 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, = 119.9

Crystals from G. Montoya, CNIO, Madrid

SAD data collection: meganuclease I-Dmol D. Mobilis

Solve a structure using MAD at Se-K edge

Fluorescence scan widget


• 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)

Human Rhinovirus 2

Crystals from N. Verdaguer, IBMB, Barcelona

Solve a structure with a unit cell par. a>400 Å


Detectors at the Experimental Hutch

Pixel main detector

Fluorescence Si-drift detector

Diamond X-ray position monitors

PIN Diodesand YAG fluorescence

screens


• 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

Area 431 x 448 mm2

Pixel size 172 mPoint spread 1 pixel

Max. framing rate 12 Hz

Dynamic Range 20 bits (106 ct)

Usable count rate 2x106 cts/s

Energy trimming Yes, recommended E0/2Gain 3, Fast, standard, high

ALBA turn = 893 ns

124 ns

Detector


• 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

Area 431 x 448 mm2

Pixel size 172 mPoint spread 1 pixel

Max. framing rate 12 Hz

Dynamic Range 20 bits (106 ct)

Usable count rate 2x106 cts/s

Energy trimming Yes, recommended E0/2Gain 3, Fast, standard, high

ALBA turn = 893 ns

124 ns

Detector. What else?

?

?


• Transmissive PIN diode• Collaboration with CNM• 10x10mm2• transmissive (10 um thick)

Other projects

• Pinhole• Heavy Z material, 80 um thick• Hole 3-20 um diameter

• Fluorescence dot• Mounted in a sample holder• 1 um diameter• Used to position beam and

spindle

• High precision tools• Sharp tips <1um• Slit blades


Thank you for your attention

The XALOC Team

First data setApril 2012

First Official Users 18 July 2012

EngineeringCarles Colldelram Mech EngineerAlex Enrique Mech TechnicianFabien Rey AlignmentMarcos Quispe Cryogenics

ComputingGuifré Cuní Controls EngineerOscar Matilla Electronic EngineerJulio LidónXavi Fariña Elec technician

ExperimentsJordi JuanhuixJordi BenachFernando GilJ.C. Martínez metrologyJosep Nicolás mirrors, metrologyIgors Sics diagnostics