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BUILDING BUILDING A USER-FRIENDLY BEAMLINE Aina Cohen and Paul Ellis.

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Presentation on theme: "BUILDING BUILDING A USER-FRIENDLY BEAMLINE Aina Cohen and Paul Ellis."— Presentation transcript:

1 BUILDING BUILDING A USER-FRIENDLY BEAMLINE Aina Cohen and Paul Ellis

2 Ideal Beamline

3 SSRL BL9-2

4 BL9-2 Oversubscribed

5 What Else Do We Have? BL 7-1 BL 1-5 BL 11-1 BL 9-2 BL 9-1

6 Beam Line 9-1 & Beam line 11-1 + Good flux + Access to useful energy ranges - 15 minutes to 1/2 hour at best to change energy ? Energy resolution

7 The Challenge: Automated Energy Moves at Side Stations To change energy at BL9-1 or BL11-1 the following must be repositioned: monochromator theta table slide (theta) monochromator bend table vertical table pitch table horizontal table yaw Weight (kg) Q315 detector: ~320 Positioners: ~550 Goniometer: ~70 Robotic Mounting System: ~90 Counter Weight: 72 Other Devices: ~45 Tabletop: 225 Total - Over 1300 kg (~3000 lbs)

8 Energy Tracking Requirements: To change energy from 12500 eV to 16500 eV, the experimental table at BL9-1 must move almost a meter (as measured from the end of the table). The mechanical components must be highly reproducible (better than 50 µm). Most of the effort to implement this system was in trouble-shooting and replacing components that were not to spec. Reliable Computer Controlled Positioners

9 Energy Tracking Requirements: Advanced Hardware Control System (DCSS) + Has access to real motors and low level resources + Passes requests from a user-friendly multi-session GUI, BLU-ICE, for data collection and staff operations + Enables pseudo-motors and a scripting engine (tcl/tk) for building complex devices Beam Line Optics Experimental Hardware Detector System Fl. Detector Sensor A/D GalilGalil NTVMS GalilGalil GalilGalil GalilGalil GalilGalil Distributed Central Server Software (DCSS) BLU-ICE GUI SGI BLU-ICE GUI SGI BLU-ICE GUI remote

10 Fit these values to a polynomial function of monochromator theta. Creating the DCSS script Optimize the beam line at different energies and record the motor positions Table Slide Position (mm) verses Monochromator Theta Difference Between Measured and Calculated Table Slide Positions (microns) TableSlide = – 2052.826104 + MonochromatorTheta x 165.354905949 – MonochromatorTheta 2 x 0.219763590 Write a Tcl/Tk script

11 Energy tracking at 9-1 and 11-1 Br - sample scans at 9-1 v. 9-2 Energy moves at beamlines 9-1 and 11-1 are now fast and accurate (useful for SAD and MAD). 9-1: 12500-16500 eV 9-2: 6000-14500 eV 11-1: 10500-15000 eV

12 Further Automation of MAD Data Collection Reliable Computer Controlled HardwareAdvanced Hardware Control System (DCSS) +

13 The Scan Tab

14 Optimization of experimental parameters for MAD

15 The Results Many structures have been solved using MAD and SAD data at BL9-1 & 11-1. Solved from BL11-1 data: TM0828 (Ribokinase) PDB: 1O14 Cell P4 1 2 1 2 (115.9x115.9x124.9x90x90x90) 2 Wavelength MAD (Above peak and remote) 2 molecules per ASU Monomer: 8 Methionines in 319 amino acids.1 32x kd Structure solution at 3.3Å resolution Solved from BL9-1 data: TM01559 (Deoxyribose-Phosphate Aldolase) PDB: 100Y Cell P2 1 (53.8x51.8x84.x 90.0x95.3x90.0) SAD (peak) 2 chains per ASU Monomer: 16 Methionines in 260 amino acids Structure solution at 1.9 Å resolution Solved from BL11-1 data: TM0920 (Alcohol Dehydrogenase) PDB: 1J5R BL11-1 Cell P2 1 (58.2x85.4x72.2x90x96.3x90) 2 Wavelength MAD (Above peak and remote) 2 molecules per ASU Monomer: 12 Methionines in 359 amino acids Structure solution at 2.6Å resolution 11-1: 2650039 (Hypothetical protein), YOR323C (Gamma-glutamyl phosphate reductase ) 9-1: TM0207 (Hypothetical protein ), TM1560 (putative serine cycle enzyme), TM1602 (Transcriptional regulator)

16 What bottlenecks remain? Sample Mounting - Hutch access is time consuming - Crystals commonly lost due to human error - Data often not collected from the best crystal Data Collection - Detector Readout Time - Exposure Times of 10 seconds or more

17 SSRL Crystal Mounting System

18 Cassette Stores 96 Samples Mount 3 cassettes at the beam line Ship 2 cassettes inside a Taylor Wharton or MVE dry shipper Store 20 cassettes inside a Taylor Wharton HC35 storage device NdFeB ring magnet Standard Hampton pins

19 The Dispensing Dewar

20 Vertically opening gripper arms The Robot and Gripper Arms Z U θ1θ1 θ2θ2 Fingers to Hold Dumbell Magnet Tool Cryo-tong Cavity Epson ES553 Robot

21 Crystal screening tab in BLU-ICE

22 Cassette Tool Kit Supplied (A) Sample Cassette and Hampton pins (B) Alignment Jig – to aid mounting pins into cassettes (C) Transfer Handle – for handling cold cassettes (D) Magnetic Tool – to mount pins in cassette & to test pin size (E) Dewar Canister – replaces stock canister in dry shipping dewars (F) Styrofoam Spacer – keeps single cassette in place when shipping (G) Teflon Ring – to support the canister in the shipping dewar Styrofoam box holds liquid nitrogen for loading cassettes

23 Video of Robot Operation

24 View of the Robot System on 1-5, 9-1, 9-2 and 11-1 9-1 1-5 9-2 9-1 11-1 11-3

25 + 3x3 array of CCD modules + Active area of 315 mm x 315 mm + 51 micron pixel size One Second Readout Raw data will be written to a 1000- Terabyte tape storage system at the San Diego Supercomputer Center. ADSC Quantum-315 Detectors at BL9-2, BL9-1, BL11-1 & BL11-3 This readout speed is 10 times faster than the Quantum-4

26 SPEAR3 The relative intensities of the SMB crystallography beamlines (~1 Å and 0.2 mm collimation) for the current SPEAR (measured) and for SPEAR3 (estimated). Beam Line11-111-39-29-17-11-5 Relative Intensity SPEAR40X15X20X15X7XX Relative Intensity SPEAR3200X75X200X75X35X100X Wavelength Range (Å)0.82-1.20.97-0.980.62-2.10.73-0.991.08 0.77- 2.1 Energy Range (keV)10-1512.6-12.85.9-2012.5-1711.55.9-16 Detector Readout (sec)111140-9010 Detector Size (mm)315 180-345188

27 Solutions Sample Mounting with SSRL Robotic System + Screen up to 288 crystals without entering the experimental hutch + Feedback systems and calibration checks ensure reliable operation + Many crystals are quickly screened and data collected from only the best Data Collection Times Reduced

28 Where do we go from here? Automated Data collection from the best crystals Automatic structure solution Automated Beam Line Alignment and Calibration Increeased Feed back - Automated Calibration Checks… Remote Access

29 New Final Beam Conditioning System

30 The Macromolecular Crystallography Group SSRL is funded by: Department of Energy, Office of Basic Energy Sciences The Structural Molecular Biology Program is supported by: National Institutes of Health, National Center for Research Resources,Biomedical Technology Program NIH, National Institute of General Medical Sciences and by the Department of Energy, Office of Biological and Environmental Research. SSRL Director Keith Hodgson SMB Leader Britt Hedman MC Leader Mike Soltis

31 Ideal Hampton Pin Lengths for Cassette Hampton Mounted CryoLoop in a MicroTube Hampton CrystalCap Magnetic Hampton 18mm CrystalCap Copper Magnetic

32 Carrier for Two Modified ALS “Pucks” Carrier that Mounts in place of Cassette In Dispensing Dewar ALS Puck with SSRL-style Ring Magnets Inside ALS Tapered Pin

33 Cut-Away Drawing Showing 4 Pucks in Dispensing Dewar

34 Fitting Table Vertical

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