eHTPX crystallization, shipping and future

1. eHTPX crystallization,shipping and future Chris Mayo, Ian Berry, Jon Diprose & Robert Esnouf Mayo et al. (2005), Structure13, 175–182.

2. OPPF crystallization facility • High-throughput nanolitre-scale sitting-drop crystallization • Hydra 96 and 2 Cartesian MicroSys 400 • 12 minutes per 96-well plate, 50 plates/day • Plate storage system • TAP Homebase, capacity 10,000 microtitre plates • Automated hand-off to imaging system • Imaging system • Veeco Oasis 1700 • ~50,000 images/day, >31,000,000 images so far • Custom IT infrastructure • Control software, Linux farm • Web-based user interface

8. A PiMS crystallization module • Module to cover management of crystallization trials • Shared development withBioXHIT allowing externalresources to contribute toPiMS goals • A self contained and well understood module • One of most demanding interms of database use anduser interface performance • Of immediate use to several PiMS sites • Four-way grouping: OPPF, NKI, EMBL Grenoble, Paris-Sud

9. Oxford eHTPX trip 28/30 Nov 2005 • 3rd eHTPX test trip • To collect some new/live data! • For testing and debugging beamline automation • To get more user experience with the sample changer • To test automated submission of frozen samples and develop interface further • To demonstrate ISPyB including remote access • For Ian and Ludovic to work on crystal/data link

10. OPPF2857 • Day 1 screened 7 crystals, nasty spots and multiple lattices but strong diffraction. Collected slightly weaker but better looking: • Spacegroup P6? • a=91.3Å, b=91.3Å, c=246.5Å • Data to 2.1Å: • ~80% complete • R= >30%

11. OPPF2857 • Day 2 screened 4 crystals refining stepwise glycerol treatment: • Spacegroup P21 • a=74.7Å, b=73.3Å, c=76.7Å, β=119.2 • All data to 1.8Å: • 91.0% complete • R= 5.5% • I/σ(I)=28.1 • Outer shell: • 71.3% complete • (98.8% at 1.94Å) • R = 53.1% • I/σ(I)=1.3

13. Feedback from 3rd eHTPX trial • Sample changer is revolutionary • Enables better science • Less agonising over marginal (3–4Å) cases • Allows a more rational approach to cryo-protection • Allows planning of data collection runs to reduce stress • Barcoding needs to be more robust • Checking could still be better and interface easier • Limitation • Unlikely to trust auto-centering for best samples • Remounting accurately would be good

14. Feedback from 3rd eHTPX trial • For Oxford the eHTPX hub was of marginal benefit • Can enter samples direct into ISPyB • Requires porting data from OPPF crystallization databases to its tray manager • Requires laborious data entry in a fixed order • Offers no mechanism for recording history of synchrotron use • Not enough flexibility for recording all forms of cryo-treatment • Pfizer group spent 6 man hours entering data already in an Excel spreadsheet! • Suggested way forward • Splitting tray manager section and crystal shipping section • More flexible and tolerant model for crystal tracking • Handling storage Dewars at home laboratories • Importing .XLS (and .XML) files directly

15. Developing a generalized container model for sample tracking / shipping • Have container description as one entry • Have instance with ‘where’, ‘valid from’ and ‘valid to’ dates • These are updated never deleted • Gives full history of all use • Could archive old records • Location • Sample changer? • Storage Dewar • Shipping Dewar • Pin (extra level for pucks) • Crystal Crystal on beam line? P7 P1 O+P4 O1 O+P6 P0 O+P5 P2 O+P3 BM14 OPPF Pfizer In transit O2 O7

16. Container model requires hierarchy