Microcrystallization System

A Microcrystallization robot was created by Berkeley Lab engineers in collaboration with scientists from the Novartis Institute of Functional Genomics. This automation can pursue thousands of crystallization conditions at the same time, thus helping to alleviate this bottleneck to the structure determination of large numbers of proteins.

This collaborative effort of researchers from Berkeley Lab, the Novartis Institute of Functional Genomics, Agencor Inc. and the Scripps Research Institute continues and builds on an FY1998/1999 project to design and develop a high throughput protein microcrystallization robotics system, called RoboHutch. This first phase of automation was highly successful. Two robotics workstations were created to automate the preparation of coarse and fine crystallization screens at the rate of about 11,500 trials per day per workstation. This represented an increase in order of magnitude in throughput of crystallization trials.

An important feature of the system was the use of precision dispensing and pipetting technology to facilitate a dramatic reduction in the volume of purified protein required for the trials. This volume was reduced from 2 microliters, a routine quantity, to the nanoliter range. Protein crystals of suitable size were successfully grown for diffraction using the nanoliter droplets.

The next phase is to further develop and test an automated system for the loading and recovery of protein microcrystals undergoing single crystal x-ray diffraction analysis. The testing will be carried out at the MCF. Specific subtasks in the project include the redesign of cryo-vial/cryo-loop assembles for the robust handling of crystals in a cryogenic environment. All manipulations are now done manually and samples can be easily damaged and become unusable. This procedure typically takes 5-15 minutes per crystal sample depending on the experience of the user. Additional time is lost moving in and out of the hutch with radiation safety protocols.

The objective is the remote and, eventually, unattended mounting and alignment of a large number of crystals in a serial processing format. Current plans call for the development of a cryogenically cooled cassette that contains multiple crystals. The crystals are mounted in place by a robot. Multiple cassettes can be daisy chained to extend the time interval of remote operation.

The project will also design a cassette manipulator for the loading and unloading of several samples in a single run at the MCF 5.0.3 beamline, develop a remotely operated protein crystal alignment system, and adapt the hardware to the alignment of the microcrystals.

For more information about the microcrystallization system, click here.