Integrated AC electrokinetic cell separation in a closed-loop device.

We integrate electrothermally induced micro-pumps and dielectrophoretic (DEP) traps into micro-circulating fluidic channel loops for yeast cell concentration and separation, two important on-chip cell manipulation tasks, with the same embedded electrodes on-chip. Each fluidic loop design contains well-defined high and low field regions that serve for both fluid transport and cellular manipulation. From a detailed study into the frequency dependent DEP behavior of viable (live) and non-viable (dead) yeast, we demonstrate several operating modes that utilize positive DEP (pDEP) and negative DEP (nDEP) to concentrate both types of cells at either the high or low electric field region and to separate one cell type to a high-field region and one to a low-field region. Because the cells visit the trapping regions repeatedly with the circulating loop design and because of the high shear rates at these stations, our device offers very rapid cell separation and concentration. Two circulating loop designs--one a four-sided square loop, the other a three-sided triangle, with different spatial symmetries and with linear dimensions less than 1 mm, are presented.

[1]  Thomas Braschler,et al.  Focusing and Continuous Separation of Cells in a Microfluidic Device using Lateral Dielectrophoresis , 2007 .

[2]  Mark A. Shannon,et al.  Sub-micrometer solid-state adhesive bonding with aromatic thermosetting copolyesters for the assembly of polyimide membranes in silicon-based devices , 2001 .

[3]  Juan G. Santiago,et al.  A planar electroosmotic micropump , 2002 .

[4]  R. Pethig,et al.  Separation of viable and non-viable yeast using dielectrophoresis. , 1994, Journal of biotechnology.

[5]  N. Aubry,et al.  Dielectrophoresis induced clustering regimes of viable yeast cells , 2005, Electrophoresis.

[6]  Ping Wang,et al.  Electrokinetic micropump and micromixer design based on ac faradaic polarization , 2004 .

[7]  H Morgan,et al.  Dielectrophoretic separation of nano-particles , 1997 .

[8]  A. Wolff,et al.  Sample preparation by cell guiding using negative dielectrophoresis , 2007 .

[9]  W. Deen Analysis Of Transport Phenomena , 1998 .

[10]  C. H. Kua,et al.  Dynamic cell fractionation and transportation using moving dielectrophoresis. , 2007, Analytical chemistry.

[11]  H. Morgan,et al.  Ac electrokinetics: a review of forces in microelectrode structures , 1998 .

[12]  Zachary Gagnon,et al.  Glutaraldehyde enhanced dielectrophoretic yeast cell separation. , 2009, Biomicrofluidics.

[13]  Thomas B. Jones,et al.  Electromechanics of Particles , 1995 .

[14]  Thomas Braschler,et al.  Continuous separation of cells by balanced dielectrophoretic forces at multiple frequencies. , 2008, Lab on a chip.

[15]  H. Morgan,et al.  Pumping of liquids with ac voltages applied to asymmetric pairs of microelectrodes. , 2003, Physical review. E, Statistical, nonlinear, and soft matter physics.

[16]  Michael P. Hughes,et al.  Nanoelectromechanics in Engineering and Biology , 2002 .

[17]  U. Zimmermann,et al.  Dielectric properties of zwitterion solutions. , 1993, Biochemical Society transactions.

[18]  Castellanos,et al.  Fluid flow induced by nonuniform ac electric fields in electrolytes on microelectrodes. I. Experimental measurements , 2000, Physical review. E, Statistical physics, plasmas, fluids, and related interdisciplinary topics.

[19]  Hsueh-Chia Chang,et al.  Electrothermal ac electro-osmosis , 2009 .

[20]  W. M. Arnold,et al.  Positioning and levitation media for the separation of biological cells , 1999, Conference Record of the 1999 IEEE Industry Applications Conference. Thirty-Forth IAS Annual Meeting (Cat. No.99CH36370).

[21]  I. Lamprecht,et al.  Dielectric properties of yeast cells as determined by electrorotation. , 1992, Biochimica et biophysica acta.

[22]  Joe S. Crane,et al.  A Study of Living and Dead Yeast Cells Using Dielectrophoresis , 1968 .

[23]  Hywel Morgan,et al.  AC ELECTROKINETICS: COLLOIDS AND NANOPARTICLES. , 2002 .

[24]  Castellanos,et al.  Fluid flow induced by nonuniform ac electric fields in electrolytes on microelectrodes. II. A linear double-layer analysis , 2000, Physical review. E, Statistical physics, plasmas, fluids, and related interdisciplinary topics.

[25]  Hywel Morgan,et al.  Dielectrophoretic separation of nano-particles , 1997 .

[26]  Zachary Gagnon,et al.  Dielectrophoretic discrimination of bovine red blood cell starvation age by buffer selection and membrane cross-linking. , 2007, Biomicrofluidics.

[27]  Jiehong Wu,et al.  Micropumping of biofluids by alternating current electrothermal effects , 2007 .

[28]  Y. Huang,et al.  Differences in the AC electrodynamics of viable and non-viable yeast cells determined through combined dielectrophoresis and electrorotation studies. , 1992, Physics in medicine and biology.