Microfluidic chips for the crystallization of biomacromolecules by counter-diffusion and on-chip crystal X-ray analysis

• Published in: Lab on a chip Vol. 9; no. 10; pp. 1412 - 1421
• Main Authors: Dhouib, Kaouthar, Khan Malek, Chantal, Pfleging, Wilhelm, Gauthier-Manuel, Bernard, Duffait, Roland, Thuillier, Gaël, Ferrigno, Rosaria, Jacquamet, Lilian, Ohana, Jeremy, Ferrer, Jean-Luc, Théobald-Dietrich, Anne, Giegé, Richard, Lorber, Bernard, Sauter, Claude
• Format: Journal Article
• Language: English
• Published: England Royal Society of Chemistry 21.05.2009
• Subjects: Biochemistry, Molecular Biology; Cellular Biology; Crystallization; Crystallography, X-Ray; Crystallography, X-Ray - instrumentation; Dimethylpolysiloxanes; Dimethylpolysiloxanes - chemistry; Life Sciences; Macromolecular Substances; Macromolecular Substances - chemistry; Microfluidic Analytical Techniques; Microfluidic Analytical Techniques - instrumentation; Molecular biology; Polymethyl Methacrylate; Polymethyl Methacrylate - chemistry
• ISSN: 1473-0197; 1473-0189
• DOI: 10.1039/b819362b

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.