Welcome!

1. Welcome! X-Ray Facility, 410-413 Kasha Laboratory Institute of Molecular Biophysics Florida State University Tallahassee, FL 32306-4380 www.sb.fsu.edu/~xray ã Thayumanasamy Somasundaram, 2003-2004

2. X-Ray Facility Orientation • History of the Facility • Facility Resources • Requirements • Data Collection • Data Processing & Manuals • Archiving & Summary • Contact & Information ã Thayumanasamy Somasundaram, 2003-2004

3. History of the Facility-1 Important Dates • Core facility planned in 1990-92 • Facility Director hired Feb 1993 • Construction June – Sep 1993 • Two generators & Image Plate (IP) detector in January 1994 • Elliott generator in 1995 ã Thayumanasamy Somasundaram, 2003-2004

4. History of the Facility-2 Important Dates • Cryo capabilities in 1996 • Crystal chamber in 1999 • Confocal mirror in Spring 2000 • CCD w/ confocal mirror in Fall 2000 • CryoCooler 700 w/ auto-refill in Fall 2000 • Additional Linux machines in 2000 • Complete renovation during 2002-03 ã Thayumanasamy Somasundaram, 2003-2004

5. Facility Resources & Accessories • Three Copper rotating anode generators • Two automated area detectors • Two confocal & one total reflection mirrors • Two cryo coolers • One automated liquid Nitrogen refiller • Linux and Windows NT computers • Full time Faculty/Staff (State-line) ã Thayumanasamy Somasundaram, 2003-2004

6. Resources-1 • Generators Two Rigaku H2R 5.4 kW generators One Elliott GX-30 3.0 kW generator • Area detector-1 (IP) R-Axis IIc (1994) w/ confocal blue mirror 65-450 mm sample-detector distance 0-25 degree non-zero 2-theta capability Oxford Cryostream 600 cold head ã Thayumanasamy Somasundaram, 2003-2004

7. Resources-2 • Area detector-2 (CCD) Mar165 CCD w/ confocal purple mirror 35-380 mm sample-CCD distance Limited non-zero 2-theta capability Oxford Cryostream 700 cold head • Long & short Supper (total-reflection) mirrors • Stand-alone IP Bio Unit I Analytical Lab ã Thayumanasamy Somasundaram, 2003-2004

8. Accessories-1 • Leica stereo microscope w/ Moticam digital camera • Nikon Optiphot microscope w/ analog camera • Stereo zoom microscopes • DVD-R & +R writers • DDS-3 & -4 tape drives • FireWire (Maxtor: 60 & 80 GB) drives • Windows NT and five Linux computers ã Thayumanasamy Somasundaram, 2003-2004

9. Accessories-2 • Cold room for 4°C crystallization set-up • Room for ambient crystallization set-up • Crystallization chamber • Temperature & humidity monitoring • Dry Cryo Shipper • Long-term sample storage • Laptop for synchrotron trips ã Thayumanasamy Somasundaram, 2003-2004

10. Requirements • Member of Structural Biology Program • Radiation safety training • Computer account (UNIX/LINUX/NT) • Body and/or finger radiation badge • No cost to use • Users bring consumables: CryoLoops, Capillaries, Tape and DVD media ã Thayumanasamy Somasundaram, 2003-2004

11. Data Collection-1 • Sample mounting decision Capillary or loop mounting? • Data collection temperature Ambient or cryo? • If cryo, cryo protection selection Grow or soak crystal? • Detector decision Image plate or CCD detector? ã Thayumanasamy Somasundaram, 2003-2004

12. Data Collection-2 • Screening or regular data collection? • Improvement or brand-new crystal? • Resolution decision Low, moderate, or high-resolution? • 2-theta angle • Cell dimension/detector distance? ã Thayumanasamy Somasundaram, 2003-2004

13. Data Collection Procedure • Check the schedule at: www.sb.fsu.edu/~soma/XraySchedules/schfrmset.html • Request time by e-mail: soma@sb.fsu.edu by phone: 644-6448 • Priority given for samples in the beam • Priority given to unstable samples • Flexible schedule & co-operation is valued ã Thayumanasamy Somasundaram, 2003-2004

14. Data Processing Procedure • Collect preliminary data & auto index • If successful, complete data collection If not, modify procedure and try • Index, integrate, scale full data set • Repeat integration & scaling, if needed • Validate and compare • Archive & transfer files ã Thayumanasamy Somasundaram, 2003-2004

15. Archiving • Data is the responsibility of the “USER” • Available archiving resources: DDS-3 & DDS-4 tape drives DVD-R & +R writers • Users bring the media • Facility archives data (last resort) • Computer Resources may be helpful ã Thayumanasamy Somasundaram, 2003-2004

16. Manuals & Documents • Online version at http://www.sb.fsu.edu/~xray/Manuals/soft_man.html • Paper version in 410 KLB (XRF) • Program authors’ website • Manufacturer’s website • IMB resource pages • Lab protocols and procedures ã Thayumanasamy Somasundaram, 2003-2004

17. Summary • State of the art facility • Excellent scientific/support staff: Computer Resources, Protein Expression Facility, Physical Biochem Facility, Electronics shop, & Machine shop • Keep it clean and in working order • Report problems to Soma (4-6448) ã Thayumanasamy Somasundaram, 2003-2004

18. Contacts & Information-1 • Data collection, processing, scheduling: Thayumanasamy Somasundaram [SOMA] Ph: 644-6448 | E-mail: soma@sb.fsu.edu www.sb.fsu.edu/~soma | www.sb.fsu.edu/~xray • XRF (KLB 410-413) | 645-1333 • Computer related: Mike Zawrotny Ph: 644-0069 | zawrotny@sb.fsu.edu ã Thayumanasamy Somasundaram, 2003-2004

19. Contacts & Information-2 • Electronics: Rick Wang Ph: 645-3309 | wang@sb.fsu.edu • Machine Shop | Wood shop: Wade Baggett or Ed Kirkland Ph: 644-1419 | 645-1338 ã Thayumanasamy Somasundaram, 2003-2004

20. Picture Gallery IP & CCD Detectors IMB (2001) ã Thayumanasamy Somasundaram, 2003-2004

21. Picture Gallery IP & CCD Detectors DLC (2002) ã Thayumanasamy Somasundaram, 2003-2004

22. Picture Gallery IP & CCD Detectors KLB (2004) ã Thayumanasamy Somasundaram, 2003-2004

23. Picture Gallery Crystal Room & Computer Room (2004) ã Thayumanasamy Somasundaram, 2003-2004