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Fourier Analysis To Measure Diffusion Coefficients and Resolve Mixtures on a Continuous Electrophoresis Chip

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A. Estévez-Torres,^† C. Gosse,*^‡ T. Le Saux,^† J.-F. Allemand,^§ V. Croquette,^§ H. Berthoumieux,^† A. Lemarchand,^‖ and L. Jullien*^†

Ecole Normale Supérieure, Département de Chimie, UMR CNRS ENS Université Paris6 8640, 24, rue Lhomond, 75005 Paris, France, Laboratoire de Photonique et de Nanostructures, LPN-CNRS, Route de Nozay, 91460 Marcoussis, France, Ecole Normale Supérieure, Département de Physique, UMR CNRS ENS Université Paris6 and Paris7 8550, 24, rue Lhomond, 75005 Paris, France, and Laboratoire de Physique Théorique de la Matiere Condensée, Université Pierre et Marie Curie Paris 6, UMR CNRS 7600, 4, place Jussieu, 75252 Paris Cedex 05, France

Anal. Chem., 2007, 79 (21), pp 8222–8231

DOI: 10.1021/ac070532z

Publication Date (Web): September 25, 2007

Abstract

We report a method to measure diffusion coefficients of fluorescent solutes in the 10²−10⁶ Da molecular mass range in a glass−PDMS chip. Upon applying a permanent electric field, the solute is introduced through a narrow channel into a wide analysis chamber where it migrates along the injection axis and diffuses in two dimensions. The diffusion coefficient is extracted after 1D Fourier transform of the resulting stationary concentration pattern. Analysis is straightforward, requiring no numerical integration or velocity field simulation. The diffusion coefficients measured for fluorescein, rhodamine green-labeled oligonucleotides, and YOYO-1-stained dsDNA fragments agree with the literature values and with our own fluorescence correlation spectroscopy measurements. As shown for 151 and 1257 base pair dsDNA mixtures, the present method allows us to rely on diffusion to quantitatively characterize the nature and the composition of binary mixtures. In particular, we implement a DNA hybridization assay to illustrate the efficiency of the proposed protocol for library screening.

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This article has been cited by 2 ACS Journal articles (2 most recent appear below).

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