Cipriani Team

Previous and current research

We develop instruments and methods for optimising X-ray scattering experiments in collaboration with the Synchrotron Crystallography Team (page 93) and the ESRF Structural Biology group, as well as contributing to the development of the EMBL@PETRA-III beamlines (see page 104). Dissemination of our technology to other synchrotrons is also an important aspect of our work.

The macromolecular crystallography (MX) beamlines rely on our MD2/MD2M diffractometers, SC3 sample changers and C3D crystal centring software to process several hundred crystals per day. The EMBL@PETRA-III MX1 beamline will also be equipped with a customised version of the MD2 diffractometer. An important task for our team is the maintenance and continuous upgrade of these instruments.

After a year of use at the ESRF MX-beamlines, the HC1b crystal dehydration device has proven to be efficient for improving the diffraction quality of protein crystals and contributed to the success of challenging projects such as the MSL proteins (Cusack Group). MAX-lab and the DLS have recently acquired an HC1b, and a collaboration defines protocols for improving the dehydration of protein crystals. A new version of the dehydration system with a sample temperature control to 5 °C, HC2, is under development.

In the framework of our collaboration with EMBL Hamburg and the ESRF, an automated BioSAXS sample environment with liquid transfer robot has been developed and installed on the ESRF/EMBL ID14-3 BioSAXS beamline. Up to eight samples and associated buffers can be automatically exposed to X-rays, and the system is optimised for sample volumes down to 5 μls with temperature control of both the sample storage and exposure cell. A fully integrated software environment with the beamline has been in use since June 2009, a copy of which is currently being validated at the X33 beamline at EMBL Hamburg. The development of a high capacity version of the machine has already started.

MX demand has now entered a phase where the capacity of existing beamline robotics is insufficient. The design of sample holders strongly impacts on the number of samples that can be affordably shipped and processed, so we have started preliminary studies on a new sample holder that is more precise and compact.

Crystal harvesting is the last step that has remained resistant to automation in MX. In collaboration with the Grenoble HTX team, we are exploring a new technology to automatically harvest crystals grown in specifically designed crystallisation supports on which we have filed a patent.

Future projects and goals

The ESRF and EMBL@PETRA-III BioSAXS beamlines will both be equipped with the final version of the automated BioSAXS sample environment. Meanwhile the HC2 dehydration device will become operational and dehydration protocols made available to users.

We will start developing a vertical kappa goniometer with a sub micrometer sphere of confusion to equip the MX2 beamline of EMBL@PETRA-III, and to upgrade the EMBL/ESRF ID23-2 micro-focus beamline.We also plan to contribute in the development of the future generation of automated beamlines, playing a driving role in the definition of a future sample holder standard for frozen crystals and associated robotic tools.

Together with the HTX team, our main objective for the future is to bridge crystallisation and data collection. Beyond automating the harvesting step, we propose a fully integrated screening platform where users can bring either protein in solution or crystallisation plates and obtain X-ray data with minimal handling. The EMBL/ESRF/Indian BM14 beamline could host such a system and become a fully integrated pilot screening platform.