Imaging neuronal seal resistance on silicon chip using fluorescent voltage-sensitive dye.

The electrical sheet resistance between living cells grown on planar electronic contacts of semiconductors or metals is a crucial parameter for bioelectronic devices. It determines the strength of electrical signal transduction from cells to chips and from chips to cells. We measured the sheet resistance by applying AC voltage to oxidized silicon chips and by imaging the voltage change across the attached cell membrane with a fluorescent voltage-sensitive dye. The phase map of voltage change was fitted with a planar core-coat conductor model using the sheet resistance as a free parameter. For nerve cells from rat brain on polylysine as well as for HEK293 cells and MDCK cells on fibronectin we find a similar sheet resistance of 10 MOmega. Taking into account the independently measured distance of 50 nm between chip and membrane for these cells, we obtain a specific resistance of 50 Omegacm that is indistinguishable from bulk electrolyte. On the other hand, the sheet resistance for erythrocytes on polylysine is far higher, at approximately 1.5 GOmega. Considering the distance of 10 nm, the specific resistance in the narrow cleft is enhanced to 1500 Omegacm. We find this novel optical method to be a convenient tool to optimize the interface between cells and chips for bioelectronic devices.

[1]  Chun-Min Lo,et al.  Impedance analysis of fibroblastic cell layers measured by electric cell-substrate impedance sensing , 1998 .

[2]  Stefano Vassanelli,et al.  Transistor Probes Local Potassium Conductances in the Adhesion Region of Cultured Rat Hippocampal Neurons , 1999, The Journal of Neuroscience.

[3]  M. Ikeda,et al.  Reflectance of rat brain structures mapped by an optical fiber technique , 1980, Journal of Neuroscience Methods.

[4]  Bernd Kuhn,et al.  Anellated hemicyanine dyes in a neuron membrane: Molecular Stark effect and optical voltage recording , 2003 .

[5]  H. Passow,et al.  Preparation and properties of human erythrocyte ghosts , 1973, Molecular and Cellular Biochemistry.

[6]  P. Fromherz,et al.  Fluorescence Interferometry of Neuronal Cell Adhesion on Microstructured Silicon , 1998 .

[7]  P. Fromherz,et al.  A neuron-silicon junction: a Retzius cell of the leech on an insulated-gate field-effect transistor. , 1991, Science.

[8]  C. Lo,et al.  Impedance analysis of MDCK cells measured by electric cell-substrate impedance sensing. , 1995, Biophysical journal.

[9]  G. Gross Simultaneous Single Unit Recording in vitro with a Photoetched Laser Deinsulated Gold Multimicroelectrode Surface , 1979, IEEE Transactions on Biomedical Engineering.

[10]  L M Loew,et al.  Spectra, membrane binding, and potentiometric responses of new charge shift probes. , 1985, Biochemistry.

[11]  W. Regehr,et al.  A long-term in vitro silicon-based microelectrode-neuron connection , 1988, IEEE Transactions on Biomedical Engineering.

[12]  P. Fromherz,et al.  Fluorescence interference-contrast microscopy of cell adhesion on oxidized silicon , 1997 .

[13]  Peter Fromherz,et al.  FREQUENCY DEPENDENT SIGNAL TRANSFER IN NEURON TRANSISTORS , 1997 .

[14]  P. Fromherz,et al.  Fast voltage transients in capacitive silicon-to-cell stimulation detected with a luminescent molecular electronic probe. , 2001, Physical review letters.

[15]  P. Fromherz,et al.  Silicon-Neuron Junction: Capacitive Stimulation of an Individual Neuron on a Silicon Chip. , 1995, Physical review letters.

[16]  P. Fromherz,et al.  Voltage-sensitive fluorescence of amphiphilic hemicyanine dyes in neuron membrane. , 1993, Biochimica et biophysica acta.

[17]  H. Ahammer,et al.  Optical multisite monitoring of cell excitation phenomena in isolated cardiomyocytes , 1995, Pflügers Archiv.

[18]  L B Cohen,et al.  Optical measurement of membrane potential. , 1978, Reviews of physiology, biochemistry and pharmacology.

[19]  Yan Zhang,et al.  Identification of endogenous outward currents in the human embryonic kidney (HEK 293) cell line , 1998, Journal of Neuroscience Methods.

[20]  Armin Lambacher,et al.  Luminescence of dye molecules on oxidized silicon and fluorescence interference contrast microscopy of biomembranes , 2002 .

[21]  P. Fromherz,et al.  Extracellular Resistance in Cell Adhesion Measured with a Transistor Probe , 2000 .

[22]  H. Itoh,et al.  Membrane conductance of an electroporated cell analyzed by submicrosecond imaging of transmembrane potential. , 1991, Biophysical journal.

[23]  Armin Lambacher,et al.  Fluorescence interference-contrast microscopy on oxidized silicon using a monomolecular dye layer , 1996 .

[24]  P. Fromherz,et al.  TIME-RESOLVED FLUORESCENCE OF A HEMICYANINE DYE : DYNAMICS OF ROTAMERISM AND RESOLVATION , 1996 .

[25]  P. Fromherz,et al.  Noninvasive neuroelectronic interfacing with synaptically connected snail neurons immobilized on a semiconductor chip , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[26]  I. Giaever,et al.  Micromotion of mammalian cells measured electrically. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[27]  B M Salzberg,et al.  Multiple site optical recording of transmembrane voltage (MSORTV) in patterned growth heart cell cultures: assessing electrical behavior, with microsecond resolution, on a cellular and subcellular scale. , 1994, Biophysical journal.

[28]  H. Feuerstein,et al.  Wie dick ist die Glykokalyx des menschlichen Erythrocyten , 1991 .

[29]  J. Pine Recording action potentials from cultured neurons with extracellular microcircuit electrodes , 1980, Journal of Neuroscience Methods.

[30]  P. Fromherz,et al.  Fluorescence of amphiphilic hemicyanine dyes without free double bonds , 1993 .

[31]  J. Westwater,et al.  The Mathematics of Diffusion. , 1957 .

[32]  A. Grinvald,et al.  Fluorescence monitoring of electrical responses from small neurons and their processes. , 1983, Biophysical journal.

[33]  Weis,et al.  Neuron adhesion on a silicon chip probed by an array of field-effect transistors. , 1996, Physical review letters.

[34]  P. Fromherz,et al.  Cable Properties of Dendrites in Hippocampal Neurons of the Rat Mapped by a Voltage‐sensitive Dye , 1997, The European journal of neuroscience.

[35]  Joachim Wegener,et al.  Barrier function of porcine choroid plexus epithelial cells is modulated by cAMP-dependent pathways in vitro , 2000, Brain Research.

[36]  Peter Fromherz,et al.  Recombinant maxi-K channels on transistor, a prototype of iono-electronic interfacing , 2001, Nature Biotechnology.

[37]  F. Graham,et al.  Characteristics of a human cell line transformed by DNA from human adenovirus type 5. , 1977, The Journal of general virology.

[38]  K. A. Jackson Mathematics of Diffusion , 2005 .

[39]  W. Maxwell Cowan,et al.  Rat hippocampal neurons in dispersed cell culture , 1977, Brain Research.

[40]  C. Pilgrim,et al.  [How thick is the glycocalyx of human erythrocytes?]. , 1991, Acta histochemica.

[41]  W. Webb,et al.  Optical imaging of cell membrane potential changes induced by applied electric fields. , 1986, Biophysical journal.

[42]  G. Loeb,et al.  A miniature microelectrode array to monitor the bioelectric activity of cultured cells. , 1972, Experimental cell research.

[43]  R. Benz,et al.  Reversible electrical breakdown of lipid bilayer membranes: A charge-pulse relaxation study , 1979, The Journal of Membrane Biology.

[44]  Peter Fromherz,et al.  Sheet conductor model of brain slices for stimulation and recording with planar electronic contacts , 2002, European Biophysics Journal.