EMBL Grenoble’s High Throughput Crystallization and Fragment Screening Facility (HTX lab), offers access to fully automated, remote controlled crystallography pipelines based on the CrystalDirect technology and the CRIMS software.
Currently The HTX Lab offers access to the following pipelines:
X-ray diffraction data collection is typically carried out at the Macromolecular crystallography (MX) beamlines of the European Synchrotron Radiation Facility (ESRF) operated by the ESRF- EMBL Joint Structural Biology Group and at the Petra III synchrotron.
In collaboration with the Synchrotron Diffraction, Instrumentation and ESRF MX Teams, the HTX lab is working towards integrating the CrystalDirect technology into the MASSIF-1 beam line environment, which will now be operated in coordination with the HTX Lab. This will increase our throughput and capabilities to support fragment screening and online crystallography. At the same time, it will enable new experimental opportunities.
Moreover, we will continue to develop the CrystalDirect technology to support serial crystallography applications as well as new approaches for the rapid analysis of membrane proteins both at cryogenic and room temperature.
For applications please contact José Marquez, head of the Crystallisation Facility.
The EMBL Grenoble HTX Lab is equipped with a nanovolume crystallization robot (including an LCP module that enables crystallization of both soluble and membrane proteins), an Echo Labcyte acoustic dispensing system, for the delivery of ligand solutions, crystal farms (at 5 º and 20 º), two robotic liquid handling stations to prepare crystallization screenings, and two automated CrystalDirect crystal harvesters with capacity to produce and store up to 400 frozen crystals per operation cycle.
The facility is operated by dedicated technicians. In the last years, and in a collaboration with the Global Phasing consortium, the facility has introduced capabilities for in line crystallographic data processing. Dedicated data storage and processing servers with capacity for 150 Tb and 84 CPUs respectively have been implemented to support data analysis.
Our services include: primary crystallization screening, crystal optimisation, automated crystal harvesting and processing and data collection in collaboration with the ESRF and PETRA III synchrotrons, as well as support for data analysis. Samples are sent by users to the facility by express courier and processed by dedicated operators at the HTX lab. Results together with all experimental parameters are made available to users in real time through our web-based CRystallization Information Management System (CRIMS). All samples received are assayed through a specifically developed method that enables to estimate the crystallization likelihood of the sample. Based on the results of this assay specific recommendations are made to users concerning the optimal incubation temperature and possible sample optimisation experiments.
In addition to the standard protocols we have developed specific procedures to handle samples with special requirements like membrane proteins for example. We are also actively involved in the development and evaluation of new technologies in macromolecular crystallization.
The HTX lab can also offer support for structure determination on a collaborative basis.
CrystalDirect technology: Cipriani & Marquez 2012 Acta Cryst. D68, 1393--1399; Zander et al. (2016) Acta Cryst. D72, 454-466
Methods for sample evaluation and quality control: Dupeux et al., 2011. Acta Cryst. D67,Mariaule et al., 2014Methods in Molecular Biology1091
On the crystallization likelihood of the sample: Dupeux et al., 2011. Acta Cryst. D67
From TNA projects: Malpartida et al. 2017. Nat Comm. 8; Bezerra et al., Sci Rep. 2017.; Wagner et al., 2017. Ange Chem 56.