157 articles – 193 references  [version française]
Detailed view Article in peer-reviewed journal
Lab on a Chip 9, 10 (2009) 1412-21
Microfluidic chips for the crystallization of biomacromolecules by counter-diffusion and on-chip crystal X-ray analysis.
Kaouthar Dhouib1, Chantal Khan Malek2, Wilhelm Pfleging3, Bernard Gauthier-Manuel2, Roland Duffait4, Gaël Thuillier2, Rosaria Ferrigno5, Lilian Jacquamet6, Jeremy Ohana6, Jean-Luc Ferrer6, Anne Théobald-Dietrich1, Richard Giegé1, Bernard Lorber1, Claude Sauter1

Microfluidic devices were designed to perform on micromoles of biological macromolecules and viruses the search and the optimization of crystallization conditions by counter-diffusion, as well as the on-chip analysis of crystals by X-ray diffraction. Chips composed of microchannels were fabricated in poly-dimethylsiloxane (PDMS), poly-methyl-methacrylate (PMMA) and cyclo-olefin-copolymer (COC) by three distinct methods, namely replica casting, laser ablation and hot embossing. The geometry of the channels was chosen to ensure that crystallization occurs in a convection-free environment. The transparency of the materials is compatible with crystal growth monitoring by optical microscopy. The quality of the protein 3D structures derived from on-chip crystal analysis by X-ray diffraction using a synchrotron radiation was used to identify the most appropriate polymers. Altogether the results demonstrate that for a novel biomolecule, all steps from the initial search of crystallization conditions to X-ray diffraction data collection for 3D structure determination can be performed in a single chip.
1:  ARN - Architecture et réactivité de l'ARN
2:  FEMTO-ST - Franche-Comté Électronique Mécanique, Thermique et Optique - Sciences et Technologies
3:  FZK - Forschungszentrum Karlsruhe
4:  CTMN - Centre de Transfert des Micro et Nanotechnologies
5:  INL - Institut des nanotechnologies de Lyon - Site d'Ecully
6:  IBS - UMR 5075 - Institut de biologie structurale