CrystalDirect: a new method for automated crystal harvesting based on laser-induced photoablation of thin films.
Cipriani, F., Rower, M., Landret, C., Zander, U., Felisaz, F. & Marquez, J.A.
Acta Crystallogr D Biol Crystallogr. 2012 Oct;68(Pt 10):1393-9. Epub 2012 Sep 18.
The use of automated systems for crystallization and X-ray data collection is now widespread. However, these two steps are separated by the need to transfer crystals from crystallization supports to X-ray data-collection supports, which is a difficult manual operation. Here, a new approach is proposed called CrystalDirect (CD) which enables full automation of the crystal-harvesting process. In this approach, crystals are grown on ultrathin films in a newly designed vapour-diffusion crystallization plate and are recovered by excision of the film through laser-induced photoablation. The film pieces containing crystals are then directly attached to a pin for X-ray data collection. This new method eliminates the delicate step of `crystal fishing', thereby enabling full automation of the crystal-mounting process. Additional advantages of this approach include the absence of mechanical stress and that it facilitates handling of microcrystals. The CD crystallization plates are also suitable for in situ crystal screening with minimal X-ray background. This method could enable the operational integration of highly automated crystallization and data-collection facilities, minimizing the delay between crystal identification and diffraction measurements. It can also contribute significantly to the advancement of challenging projects that require the systematic testing of large numbers of crystals.
New beamline dedicated to solution scattering from biological macromolecules at the ESRF.
Pernot, P., Theveneau, P., Giraud, T., Nogueira Fernandes, R., Nurizzo, D., Spruce, D., Surr, J., McSweeney, S., Round, A., Felisaz, F., Foedinger, L., Gobbo, A., Huet, J., Villard, C. & Cipriani, F.
J. Phys.: Conf. Ser. 247 (2010) 012009
The new bio-SAXS beamline (ID14-3 at the ESRF, Grenoble, France) is dedicated exclusively to small-angle scattering experiments of biological macromolecules in solution and has been in user operation since November 2008. Originally a protein crystallography beamline, ID14-3 was refurbished, still as a part of the ESRF Structural Biology group, with the main aim to provide a facility with 'quick and easy' access to satisfy rapidly growing demands from crystallographers, biochemists and structural biologists. The beamline allows manual and automatic sample loading/unloading, data collection, processing (conversion of a 2D image to a normalized 1D X-ray scattering profile) and analysis. The users obtain on-line standard data concerning the size (radius of gyration, maximum dimension and volume) and molecular weight of samples which allow on-the fly ab-inito shape reconstruction in order to provide feedback enabling the data collection strategies to be optimized. Automation of sample loading is incorporated on the beamline using a device constructed in collaboration between the EMBL (Grenoble and Hamburg outstations) and the ESRF. Semi/automated data analysis is implemented following the model of the SAXS facility at X33, EMBL Hamburg. This paper describes the bio-SAXS beamline and set-up characteristics together with the examples of user data obtained.
Improving diffraction by humidity control: a novel device compatible with X-ray beamlines.
Sanchez-Weatherby, J., Bowler, M.W., Huet, J., Gobbo, A., Felisaz, F., Lavault, B., Moya, R., Kadlec, J., Ravelli, R.B. & Cipriani, F.
Acta Crystallogr D Biol Crystallogr. 2009 Dec;65(Pt 12):1237-46. Epub 2009Nov 17.
Dehydration of protein crystals is rarely used, despite being a post-crystallization method that is useful for the improvement of crystal diffraction properties, as it is difficult to reproduce and monitor. A novel device for hydration control of macromolecular crystals in a standard data-collection environment has been developed. The device delivers an air stream of precise relative humidity that can be used to alter the amount of water in macromolecular crystals. The device can be rapidly installed and is fully compatible with most standard synchrotron X-ray beamlines. Samples are mounted in cryoloops and the progress of dehydration can be monitored both optically and by the acquisition of diffraction images. Once the optimal hydration level has been obtained, cryocooling is easy to achieve by hand or by using a sample changer. The device has been thoroughly tested on several ESRF beamlines and is available to users.
Automation of sample mounting for macromolecular crystallography.
Cipriani, F., Felisaz, F., Launer, L., Aksoy, J.S., Caserotto, H., Cusack, S., Dallery, M., di-Chiaro, F., Guijarro, M., Huet, J., Larsen, S., Lentini, M., McCarthy, J., McSweeney, S., Ravelli, R., Renier, M., Taffut, C., Thompson, A., Leonard, G.A. & Walsh, M.A.
Acta Crystallogr D Biol Crystallogr. 2006 Oct;62(Pt 10):1251-9. Epub 2006Sep 19.
A standard sample holder and vial for cryocooled macromolecular crystals has been defined for use with robotic sample changers. This SPINE standard sample holder is a modified version, with added features and specifications, of sample holders in common use. In particular, the SPINE standard meets the precision required for automatic sample exchange and includes a cap that is identified by a two-dimensional datamatrix code as well as an optional vial. At the ESRF, the sample holder standard is in use with the EMBL/ESRF/BM14 robotic sample changer (SC3) which is installed on eight beamlines. The SC3 can hold up to 50 crystals stored in five baskets. A datamatrix reader in the SC3 ensures safe management of the sample flow and facilitates fully automatic screening and characterization of samples. Tools for handling and transporting 50 samples in a dry shipping dewar have been developed. In addition to the SC3, the SPINE sample holder is currently compatible with a number of other robotic sample changers.