eHTPX crystallization, shipping and future Chris Mayo, Ian Berry, Jon Diprose & Robert Esnouf Mayo et al. (2005), Structure 13, 175–182.

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

A PiMS crystallization module Module to cover management of crystallization trials Shared development with BioXHIT allowing external resources to contribute to PiMS goals A self contained and well understood module One of most demanding in terms of database use and user interface performance Of immediate use to several PiMS sites Four-way grouping: OPPF, NKI, EMBL Grenoble, Paris-Sud

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

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%

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

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

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

Developing a generalized container model for sample tracking / shipping Location Sample changer? Storage Dewar Shipping Dewar Pin (extra level for pucks) Crystal 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 Crystal on beam line? P7 P1 O+P4 O1 P0 O+P6 O+P5 P2 O+P3 BM14 OPPF Pfizer In transit O2 O7

Container model requires hierarchy