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International audience; Here, we describe a plug-and-play microfluidic platform, suitable for protein crystallization. The droplet factory is designed to generate hundreds of droplets as small as a few nanoliters (2 to 10nL) for screening and optimization of crystallization conditions. Commercially-available microfluidic junctions and tubing are combined to create the appropriate geometry. In addition, a " chemical library " is produced in tubing. The microfluidic geometry for a " crystallization agent-based chemical library " is validated by screening crystallization conditions of lysozyme. The microfluidic geometry for a " ligand-based chemical library " is also explored to co-crystallize the protein QR2 with a ligand, for the purposes of structure-based drug design. This platform mixes aqueous phases (containing the protein and the crystallization agent) and organic phases (containing the ligand), during the droplet generation and circulation without using any surfactant. The droplet composition is controlled by the respective flow-rates of the different solutions, and checked by measuring on-line absorbance. The low volumes involved in the crystallization trials, the speed of execution and the absence of a microfabrication stage make our platform a cheap, easy-to-use and versatile tool for crystallization studies.

International audience; We describe a universal, high-throughput droplet-based microfluidic platform for crystallization. It is suitable for a multitude of applications, due to its flexibility, ease of use, compatibility with all solvents and low cost. The platform offers four modular functions: droplet formation, on-line characterization, incubation and observation. We use it to generate droplet arrays with a concentration gradient in continuous long tubing, without using surfactant. We control droplet properties (size, frequency and spacing) in long tubing by using hydrodynamic empirical relations. We measure droplet chemical composition using both an off-line and a real-time on-line method. Applying this platform to a complicated chemical environment, membrane proteins, we successfully handle crystallization, suggesting that the platform is likely to perform well in other circumstances. We validate the platform for fine-gradient screening and optimization of crystallization conditions. Additional on-line detection methods may well be integrated into this platform in the future, for instance, an on-line diffraction technique. We believe this method could find applications in fields such as fluid interaction engineering, live cell study and enzyme kinetics.

A microfluidic platform was used to address the problems of obtaining diffraction-quality crystals and crystal handling during transfer to the X-ray diffractometer. Crystallization conditions of a protein of pharmaceutical interest were optimized and X-ray data were collected both in situ and ex situ.

We present a simple and easy-to-use microfluidic setup for the crystallization of mineral, organic, and biological materials. After briefly presenting the hydrodynamic properties of the setup, we test and validate it with viscous media by crystallizing a protein in an aqueous solution of PEG. We obtain nucleation data in nanocrystallizers using a droplet-based method and precisely controlling input flows to test different crystallization conditions.

This paper highlights the work of our group on the control and the observation of nucleation with techniques using nanotechnologies. This control is performed either by triggering nucleation in time with an external field or by localising it spatially in a microdroplet. Localisation in time using light irradiation induces nucleation by forming radicals; the use of electric field acts locally on the density of the solution. Localisation in space with a microfluidic device produces hundreds of nanovolume crystallisers where concentration and temperature are easily monitored. Thus, accurate statistical studies lead to the nucleation parameters (metastable zone, nucleation rate and polymorphism). Lastly, confinement with a microdroplet generator permits to reach very high supersaturations in fL to pL volumes allowing nucleation of a single crystal per microdroplet. All these methods clearly enhance nucleation in the metastable zone. Finally, they use small quantities of products offering potentialities for the screening of crystallisation conditions and phases (polymorphism).