The CIMR offers structural biologists facilities for high-throughput initial identification and optimization of crystallization conditions. This facility is maintained by Giles Lewis, who is responsible for the equipment, training and supporting users, and assisting with method development.
Once a sample has been successfully crystallized, crystals can be screened in house using an x-ray generator facility shared with the MRC’s Mitochondrial Biology Unit. Data collection is done using the in-house generator or at a synchrotron facility, most commonly the UK’s Diamond Light Source. Data collection and processing at Diamond can be controlled remotely from a terminal within the CIMR.
We have begun to develop integration with Diamond’s new in-situ x-ray diffraction facilities, allowing scientists to move directly from a successful crystallization experiment to remote data collection without requiring further sample preparation or crystal handling.
Prepared crystallization conditions. Premade 96-condition sitting drop crystallization trays spanning multiple different strategies from commercial reagent suppliers are kept in stock for standard experiments. Optimization screens based on commercial or in-house reagents are available on request.
Innovadyne ScreenMaker. This instrument dispenses sample and reagent sitting drops into standard format 96-well SBS plates, with a minimum drop volume of 100nl. Large crystallization screens can be rapidly set up with a small total sample volume compared to manual approaches.
Tecan Freedom Evo. This liquid handling system is used by the facility to prepare commercial and custom screens in a variety of plate formats.
Formulatrix Rock Imager. This system holds five hundred crystallization experiment trays. Any plate design with a standard SBS footprint can be accomodated. The trays are kept at a constant temperature and individual drops are automatically imaged according to user-defined schedule. The system’s imaging microscope is fitted with a polarizing filter to enable crystals to be examined for birefringence. Images collected are stored in a database along with details of the experiment and screening conditions, and can be analysed using Formulatrix’s Rock Maker software.
Crystal storage and mounting. The CIMR is currently fitting out a new dedicated space with equipment for scooping, mounting, and freezing crystals.
X-ray generator and detectors. Jointly with the MRC Mitochondrial Biology Unit, the CIMR maintains a Rigaku FR-E+ X-ray generator equipped with MAR 345dtb image plate detectors. This allows in-house screening of crystals and, depending on crystal quality, collection of data for structure solution.
Diamond remote terminal. A dedicated workstation configured for remote access to the Diamond Light Source’s beamline and compute facilities is available.
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