Dielectric spectroscopy in a micromachined flow cytometer: theoretical and practical considerations.

We propose a model to determine the influence of different cell properties, such as size, membrane capacitance and cytoplasm conductivity, on the impedance spectrum as measured in a microfabricated cytometer. A dielectric sphere of equivalent complex permittivity is used as a simplified model to describe a biological cell. The measurement takes place between a pair of facing microelectrodes in a microchannel filled with a saline solution. The model incorporates various cell parameters, such as dielectric properties, size and position in the channel. A 3D finite element model is used to evaluate the magnitude of the electric field in the channel and the resultant changes in charge densities at the measurement electrode boundaries as a cell flows past. The charge density is integrated on the electrode surface to determine the displacement current and the channel impedance for the computed frequency range. The complete impedance model combines the finite element model, the electrode-electrolyte interface impedance and stray impedance, which are measured from a real device. The modeled dielectric complex spectra for various cell parameters are discussed and a measurement strategy for cell discrimination with such a system is proposed. We finally discuss the amount of noise and measurement fluctuations of the sensor.

[1]  S Takashima,et al.  Frequency domain analysis of membrane capacitance of cultured cells (HeLa and myeloma) using the micropipette technique. , 1990, Biophysical journal.

[2]  Peter R. C. Gascoyne,et al.  General expressions for dielectrophoretic force and electrorotational torque derived using the Maxwell stress tensor method , 1997 .

[3]  Ulrich Zimmermann,et al.  A single-shell model for biological cells extended to account for the dielectric anisotropy of the plasma membrane , 2001 .

[4]  T. Pajkossy,et al.  Impedance of rough capacitive electrodes , 1994 .

[5]  C. P. Bean,et al.  Counting and Sizing of Submicron Particles by the Resistive Pulse Technique , 1970 .

[6]  R. Hoffman,et al.  Flow-system measurement of cell impedance properties. , 1979, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

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

[8]  Alan G. R. Evans,et al.  Design and fabrication of a micromachined Coulter counter , 1999 .

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

[10]  S. Gawad,et al.  Leukocytes Discrimination by Impedance Spectroscopy Flow Cytometry , 2002 .

[11]  J. A. Connelly,et al.  Low noise electronic system design , 1993 .

[12]  E. Prodan,et al.  The dielectric behaviour of living cell suspensions , 1999 .

[13]  A. Irimajiri,et al.  Dielectric analysis of mitochondria isolated from rat liver. I. Swollen mitoplasts as simulated by a single-shell model. , 1984, Biochimica et biophysica acta.

[14]  Towards Single-Cell-Controlled Electroporation in a Microfluidic Device , 2002 .

[15]  W. Kenan,et al.  Impedance Spectroscopy: Emphasizing Solid Materials and Systems , 1987 .

[16]  C. Cametti,et al.  Reduction of the contribution of electrode polarization effects in the radiowave dielectric measurements of highly conductive biological cell suspensions. , 2001, Bioelectrochemistry.

[17]  Roland Schinzinger,et al.  Conformal Mapping: Methods and Applications , 1991 .

[18]  J. Maxwell A Treatise on Electricity and Magnetism , 1873, Nature.

[19]  S Takashima,et al.  Dielectric properties of mouse lymphocytes and erythrocytes. , 1989, Biochimica et biophysica acta.

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

[21]  Bernhard E. Boser,et al.  Microfabricated multi-frequency particle impedance characterization system , 2000 .

[22]  Uwe Pliquett,et al.  Passive electrical properties of RBC suspensions: changes due to distribution of relaxation times in dependence on the cell volume fraction and medium conductivity , 1998 .

[23]  G. Fuhr,et al.  Dielectric spectroscopy of single human erythrocytes at physiological ionic strength: dispersion of the cytoplasm. , 1996, Biophysical journal.

[24]  A. Irimajiri,et al.  A dielectric theory of "multi-stratified shell" model with its application to a lymphoma cell. , 1979, Journal of theoretical biology.

[25]  K. Shirahige,et al.  Progression of cell cycle monitored by dielectric spectroscopy and flow‐cytometric analysis of DNA content , 2000, Yeast.

[26]  Christopher L. Davey,et al.  The influence of electrode polarisation on dielectric spectra, with special reference to capacitive biomass measurements: /II Reduction in the contribution of electrode polarisation to dielectric spectra using a two-frequency method , 1998 .

[27]  Christopher L. Davey,et al.  The influence of electrode polarisation on dielectric spectra, with special reference to capacitive biomass measurements I. Quantifying the effects on electrode polarisation of factors likely to occur during fermentations , 1998 .

[28]  S. Quake,et al.  A microfabricated fluorescence-activated cell sorter , 1999, Nature Biotechnology.

[29]  Koji Asami,et al.  Characterization of biological cells by dielectric spectroscopy , 2002 .

[30]  Alan P. Morrison,et al.  Development of a microfluidic device for fluorescence activated cell sorting , 2002 .

[31]  Yuri Feldman,et al.  Electrode polarization correction in time domain dielectric spectroscopy , 2001 .