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Acta Crystallogr Sect F Struct Biol Cryst Commun. Aug 1, 2011; 67(Pt 8): 971–975.
Published online Jul 26, 2011. doi:  10.1107/S1744309111024456
PMCID: PMC3151141
From screen to structure with a harvestable microfluidic device
Vivian Stojanoff,a Jean Jakoncic,a Deena A. Oren,b V. Nagarajan,c Jens-Christian Navarro Poulsen,d Melanie A. Adams-Cioaba,e Terese Bergfors,f and Morten O. A. Sommere*
aNational Synchrotron Light Source, Brookhaven National Laboratories, Upton, NY 11973, USA
bStructural Biology Resource Center, Rockefeller University, 1230 York Avenue, New York, NY 10065, USA
cJAN Scientific Inc., 4726 Eleventh Avenue NE, Suite 101, Seattle, WA 98105, USA
dDepartment of Chemistry, Biophysical Chemistry Group, University of Copenhagen, Universitetsparken 5, DK-2100 Copenhagen, Denmark
eMicrolytic ApS, Universitetsparken 7, DK-4000 Roskilde, Denmark
fDepartment of Cell and Molecular Biology, Uppsala University, SE-75 124 Uppsala, Sweden
Correspondence e-mail: ms/at/microlytic.com
Received March 22, 2011; Accepted June 21, 2011.
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
Articles from Acta Crystallographica Section F: Structural Biology and Crystallization Communications are provided here courtesy of
International Union of Crystallography