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Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 August 1; 67(Pt 8): 971–975.
Published online 2011 July 26. doi:  10.1107/S1744309111024456
PMCID: PMC3151141

From screen to structure with a harvestable microfluidic device

Abstract

Advances in automation have facilitated the widespread adoption of high-throughput vapour-diffusion methods for initial crystallization screening. However, for many proteins, screening thousands of crystallization conditions fails to yield crystals of sufficient quality for structural characterization. Here, the rates of crystal identification for thaumatin, catalase and myoglobin using microfluidic Crystal Former devices and sitting-drop vapour-diffusion plates are compared. It is shown that the Crystal Former results in a greater number of identified initial crystallization conditions compared with vapour diffusion. Furthermore, crystals of thaumatin and lysozyme obtained in the Crystal Former were used directly for structure determination both in situ and upon harvesting and cryocooling. On the basis of these results, a crystallization strategy is proposed that uses multiple methods with distinct kinetic trajectories through the protein phase diagram to increase the output of crystallization pipelines.

Keywords: Crystal Former, protein crystallization, structural biology, liquid–liquid diffusion, microfluidics

Les articles de Acta Crystallographica Section F: Structural Biology and Crystallization Communications ont été offerts à titre gracieux par International Union of Crystallography.