Lab Spotlight: 

The HTX Lab at EMBL Grenoble

A high-throughput, automated crystallization platform

The High Throughput Crystallization Laboratory is a major facility for automated, nanovolume, crystallization screening located at the EMBL (European Molecular Biology Laboratory) Grenoble Outstation. 

The standard plates routinely used at the HTX platform are the CrystalDirectTM 96-well plates, which are ideally suited for crystallization experiments, but also for in situ, serial data collection and LCP experiments. When used in combination with the CrystalDirectTM Harvester (developed by the Marquez & Cipriani teams), this gives access to fully automated crystal soaking, cryo-cooling and harvesting (Zander et al., 2016). By removing the bottlenecks represented by manual crystal processing, the CrystalDirectTM Technology is accelerating progress for challenging projects that require the analysis of many samples. Moreover, it precludes the need to add cryo-protectants before cryo-cooling (Bezerra et al., 2017, Fernandez et al., 2017, Zander et al., 2016). 

The HTX lab has also developed the web-based Crystallization Information Management System (CRIMS), which is currently used by multiple laboratories.  The combination of the CrystalDirectTM Technology with the ESRF MASSIF-1 as well as the CRIMS and ISPyB systems has enabled the development of fully automated, protein-to-structure pipelines, integrating crystallization, crystal mounting and processing as well as X-ray data collection and analysis in a continuous, remote-controlled workflow.

These pipelines are being applied to support challenging projects in structural biology as well as to provide access to large-scale fragment screening. Scientists working at either academic labs or industry can access the HTX platform services, offered via the EU-funded iNEXT program (in collaboration with ESRF) and EMBLEM, respectively. Furthermore, all users have remote web-based access to experimental set-up and results in real-time (Marquez & Cipriani, 2014).

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The HTX Team lead by Dr. José Márquez.
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CrystalDirect™ crystallization plate during automated crystal harvesting and cryo-cooling with the CrystalDirect™ Harvester.

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CrystalDirect® offers a large amount of cut shapes for the laser photoablation process. The different shapes adapt to different protein crystal distributions and orientations on the native membrane. Consequently, CrystalDirect® can select and harvest single crystals, batches of crystals, micro-crystals and single needles from clusters.

Recommended Tools:

"We recommend the CrystalDirect™ plates for their versatility, compatibility and ease-of-use"

MiTeGens 96-well CrystalDirect™ Plates

Contact:

DR. JOSÉ A. MÁRQUEZ
Head of Crystallisation Facility
[email protected]
https://htxlab.embl.org/

References:

Bezerra GA, Ohara-Nemoto Y, Cornaciu I, Fedosyuk S, Hoffmann G, Round A, Márquez JA, Nemoto TK, Djinović-Carugo K. (2017) Bacterial protease uses distinct thermodynamic signatures for substrate recognition. Sci Rep 7(1) doi: 10.1038/s41598-017-03220-y

Fernández I, Cornaciu I, Del Carmen Carrica M, Uchikawa E, Hoffmann G, Sieira R, Márquez JA, Goldbaum FA. (2017) Three-dimensional structure of full-length NtrX, an unusual member of the NtrC family of response regulators. J. Mol. Biol. doi: 10.1016/j.jmb.2016.12.022

Márquez JA, Cipriani F. (2014) CrystalDirect™: a novel approach for automated crystal harvesting based on photoablation of thin films Methods Mol. Biol. 1091:197-203. doi: 10.1007/978-1-62703-691-7_14

Zander U, Hoffmann G, Cornaciu I, Marquette JP, Papp G, Landret C, Seroul G, Sinoir J, Röwer M, Felisaz F, Rodriguez-Puente S, Mariaule V, Murphy P, Mathieu M, Cipriani F, Márquez JA. (2016) Automated harvesting and processing of protein crystals through laser photoablation. Acta Crystallogr D Struct Biol 72(Pt 4):454-466. doi: 10.1107/s2059798316000954