Microfluidic Cell Separation by 2-dimensional Dielectrophoresis

We describe a microfluidic device for separating cells according to their dielectric properties by combining 2-dimensional dielectrophoretic forces with field-flow-fractionation. The device comprises a thin chamber in which a travelling-wave electrical field is generated by a planar, multilayer microelectrode array at the bottom. Under the balance of gravitational and dielectrophoretic levitation forces, cells introduced into the device are positioned at different equilibrium heights in a velocity profile established inside the chamber, and thereby transported at different velocities by the fluid. Simultaneously, cells are subjected to a horizontal travelling-wave dielectrophoretic force that deflects them across the flow stream. The 2-dimensional dielectrophoretic forces acting on cells and the associated velocities in the fluid-flow and travelling-field directions depend sensitively on cell dielectric properties. The responses of cultured MDA-435 human breast cancer, HL-60 human leukemia and DS19 murine erythroleukemia cells, and of peripheral blood mononuclear (PBMN) cells were studied as functions of the frequency and voltage of the applied electric signals, and of the fluid flow rate. Significant differences were observed between the responses of different cell types. Cell separation was demonstrated by the differential redistribution of MDA-435 and PBMN cells as they flowed through the device. The device can be readily integrated with other microfluidic components for microscale sample preparation and analysis.

[1]  K. Kaler,et al.  Dielectrophoretic spectra of single cells determined by feedback-controlled levitation. , 1990, Biophysical journal.

[2]  R. Austin,et al.  Deformation and flow of red blood cells in a synthetic lattice: evidence for an active cytoskeleton. , 1995, Biophysical journal.

[3]  R. Pethig Dielectrophoresis: Using Inhomogeneous AC Electrical Fields to Separate and Manipulate Cells , 1996 .

[4]  Ulrich Zimmermann,et al.  A traveling‐wave micropump for aqueous solutions: Comparison of 1 g and μg results , 1993 .

[5]  M. Heller,et al.  Preparation and hybridization analysis of DNA/RNA from E. coli on microfabricated bioelectronic chips , 1998, Nature Biotechnology.

[6]  G. Fuhr,et al.  Three-dimensional electric field traps for manipulation of cells--calculation and experimental verification. , 1993, Biochimica et biophysica acta.

[7]  Ronald Pethig,et al.  Dielectrophoretic characterization and separation of micro-organisms , 1994 .

[8]  Y. Huang,et al.  Electrokinetic behaviour of colloidal particles in travelling electric fields: studies using yeast cells , 1993 .

[9]  Thomas Koch,et al.  Characterization of near-wall hydrodynamic lift forces using sedimentation field-flow fractionation , 1992 .

[10]  J. Hodgson,et al.  DNA chips: An array of possibilities , 1998, Nature Biotechnology.

[11]  F F Becker,et al.  Membrane changes associated with the temperature-sensitive P85gag-mos-dependent transformation of rat kidney cells as determined by dielectrophoresis and electrorotation. , 1996, Biochimica et biophysica acta.

[12]  Mann A. Shoffner,et al.  Integrated cell isolation and polymerase chain reaction analysis using silicon microfilter chambers. , 1998, Analytical biochemistry.

[13]  Y. Huang,et al.  Introducing dielectrophoresis as a new force field for field-flow fractionation. , 1997, Biophysical journal.

[14]  F F Becker,et al.  Cell separation on microfabricated electrodes using dielectrophoretic/gravitational field-flow fractionation. , 1999, Analytical chemistry.

[15]  F F Becker,et al.  Non-uniform spatial distributions of both the magnitude and phase of AC electric fields determine dielectrophoretic forces. , 1995, Biochimica et biophysica acta.

[16]  S. Masuda,et al.  Handling biological cells using a fluid integrated circuit , 1990 .

[17]  F F Becker,et al.  Dielectrophoretic manipulation of cells with spiral electrodes. , 1997, Biophysical journal.

[18]  Masahiro Iwadare,et al.  Separation of Small Particles Suspended in Liquid by Nonuniform Traveling Field , 1987, IEEE Transactions on Industry Applications.

[19]  Peter R C Gascoyne,et al.  Dielectrophoretic Separation of Cancer Cells from Blood. , 1997, IEEE transactions on industry applications.

[20]  Michael G. Ormerod,et al.  Flow Cytometry: A Practical Approach , 1994 .

[21]  Ulrich Zimmermann,et al.  Electro-rotation: development of a technique for dielectric measurements on individual cells and particles , 1988 .

[22]  Ronald Pethig,et al.  DEP-FFF: Field-Flow Fractionation Using Non-Uniform Electric Fields , 1997 .

[23]  Ronald Pethig,et al.  Selective dielectrophoretic confinement of bioparticles in potential energy wells , 1993 .

[24]  R. Pethig,et al.  Dielectrophoretic separation of cells: Continuous separation , 1995, Biotechnology and bioengineering.