W0352

A high throughput protein crystallization facility is being developed at Lawrence Livermore National Laboratory (LLNL) with the support of the TB Structural Genomics Center. The design goals were to develop a low cost, modular system that would be suitable for use in other academic laboratories. Our efforts have led to the development of one new robotic platform based on the widely used Robbins Hydra dispenser (Hydra Plus One) and a new crystallization microplate, IntelliPlate (developed in conjunction with Art Robbins Enterprises). We also worked with Velocity11 to adapt their Elispot detector into a microplate reader capable of high resolution imaging of the wells of any 96 well SBS format microplate. The Hydra Plus One is a 96 channel dispenser augmented with a single channel touchless dispenser. The touchless dispenser is used to deposit a protein drop within the range of 200nL to 2uL into the small circular well of the IntelliPlate; the 96 channel dispenser delivers a matching volume of crystallization cocktail to the drop and fills the reservoir. The Velocity11 Versascan C-type is an automation ready, high resolution CCD microscope compatible with SBS format plates. The Versascan acquires 1 Megapixel images of each drop, with an average speed of 2s per drop. The IntelliPlate is a 96 well, SBS footprint, sitting drop microplate designed for increased range of usable drop volumes (nanoliter to microliter), improved sealing, ease of crystal mounting and optical clarity. The well for the protein drop is semi-spherical which provides repeatable centering for any size drop. The optics and protein well geometry of the IntelliPlate allow for automated plate scoring by eliminating aberrations in the images caused by lighting and variable drop shape. Technologies developed through the LLNL Crystallization Facility are available and enable modular, lab scale high throughput crystallization.