# US 8,784,633 B2 · Automatic positioning and sensing microelectrode arrays > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,784,633 B2 - Document ID: US8784633B2 - Status: Granted - Kind: B2 - Application number: 12/667,507 - Filed: 2007-07-04 - Published: 2010-10-28 - Granted: 2014-07-22 - Earliest claimed: 2007-07-04 - Assignees: CapitalBio Corporation; Tsinghua University - Applicants: Guangxin Xiang; Liangbin Pan; Wanli Xing; Lihua Huang; Zhongyao Yu; Jing Zhu; Yuxiang Zhou; Jing Cheng - Inventors: Guangxin Xiang; Liangbin Pan; Wanli Xing; Lihua Huang; Zhongyao Yu; Jing Zhu; Yuxiang Zhou; Jing Cheng - Web page: https://patentgrep.com/patent/US8784633B2 ## CPC - G01N 33/4836: {using multielectrode arrays} - B01L 3/502707: {characterised by the manufacture of the container or its components} - B01J 2219/00653: the compounds being bound to electrodes embedded in or on the solid supports - B01J 2219/00743: Cells - B01L 2300/0645: Electrodes - B01L 2400/0421: electrophoretic flow - B01L 2400/0424: Dielectrophoretic forces - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE - B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS ## Abstract A microelectrode sensing device includes a substrate and an array of microelectrode sensors. Each sensor includes a first conductive layer that at least partially conducts electricity. The first conductive layer is formed above the substrate and patterned to include a recording electrode that measures electrical activities of target cells. Each sensor also includes a second conductive layer that at least partially conducts electricity. The second conductive layer is elevated above the first layer and patterned to include multiple positioning electrodes arranged to define a sensing region above the recording electrode. The positioning electrodes are designed to generate an electric field pattern in the sensing region to move and confine the target cells to a sub-region of the sensing region that at least partially overlaps the recording electrode. ## Claims ### Claim 1 A microelectrode sensing device, comprising: a substrate; and an array of microelectrode sensors formed on the substrate, each sensor comprising a first conductive layer, that at least partially conducts electricity, formed above the substrate and patterned to comprise a recording electrode to measure electrical activities of one or more target cells in a solution; a second conductive layer, that at least partially conducts electricity, elevated above the first layer and patterned to comprise a plurality of positioning electrodes arranged to define a sensing region above the recording electrode in which the solution is located, the positioning electrodes operable to generate an electric field pattern in the sensing region to move and confine the target cells to a sub-region of the sensing region that at least partially overlaps the recording electrode; and an insulator material filled between the first conductive layer and the second conductive layer to electrically insulate the recording electrode from the positioning electrodes. ### Claim 2 (depends on 1) The microelectrode sensing device of claim 1, wherein the sub-region comprises locations of minimal electrical field strength. ### Claim 3 (depends on 1) The microelectrode sensing device of claim 1, wherein the sub-region is located substantially near a center of the sensing region. ### Claim 4 (depends on 1) The microelectrode sensing device of claim 1, further comprising at least four positioning electrodes. ### Claim 5 (depends on 4) The microelectrode sensing device of claim 4, wherein four of the positioning electrodes are configured to form a first pair of electrodes and a second pair of electrodes operable to apply a first signal and a second signal to generate the electric field pattern; and expose the target cells in the sensing region to one or more dielectrophoretic forces that are generated based on the electric field pattern. ### Claim 6 (depends on 5) The microelectrode sensing device of claim 5, wherein the first and second signals comprise a pair of alternating current signals; and the one or more dielectrophoretic forces generated comprises a negative dielectrophoretic force. ### Claim 7 (depends on 6) The microelectrode sensing device of claim 6, wherein the first and second pairs of electrodes are configured to apply the negative dielectrophoretic force to confine the target cells into an ordered pattern of cells. ### Claim 8 (depends on 6) The microelectrode sensing device of claim 6, wherein the pair of alternating current signals comprises a signal with an amplitude of 2 volts and a frequency of 5 megahertz. ### Claim 9 (depends on 6) The microelectrode sensing device of claim 6, wherein the pair of alternating current signals applied are separated by a phase angle difference of 180 degrees. ### Claim 10 (depends on 1) The microelectrode sensing device of claim 1, wherein the positioning electrodes are further operable to generate electrical potentials based on a second-order polynomial that obeys Laplace's equation. ### Claim 11 (depends on 10) The microelectrode sensing device of claim 10, wherein the positioning electrodes have a shape based on equipotential boundaries determined based on the generated electrical potentials. ### Claim 12 (depends on 1) The microelectrode sensing device of claim 1, wherein the first conductive layer and the second conductive layer are arranged to reduce capacitive or inductive interference. ### Claim 13 (depends on 1) The microelectrode sensing device of claim 1, further comprising one or more passivation layers arranged to form barriers that at least partially confine the solution within the sensing region. ### Claim 14 (depends on 1) The microelectrode sensing device of claim 1, wherein the positioning electrodes are further operable to apply signals that selectively lyse one or more of the target cells. ### Claim 15 (depends on 1) The microelectrode sensing device of claim 1, wherein each of the sensors in the array is configured to move and confine the target cells independent of other sensors in the array. ### Claim 16 (depends on 1) The microelectrode sensing device of claim 1, wherein the recording electrode is further configured to apply a positive dielectrophoretic force. ### Claim 17 (depends on 1) The microelectrode sensing device of claim 1, further comprising a signal generator configured to apply one or more signals through the positioning electrodes or the recording electrode. ### Claim 18 A method comprising: providing a microelectrode sensing device including forming a recording electrode in a first conductive layer, that at least partially conducts electricity, over a substrate; arranging a plurality of positioning electrodes in a second conductive layer, that at least partially conducts electricity, over the recording electrode to define a sensing region over the recording electrode and reduce capacitive or inductive interference; adding a passivation material at least between the first and second conductive layers arranged to electrically insulate the recording electrode from the positioning electrodes and retain one or more target cells in a solution; using the positioning electrodes to apply an electric field pattern that moves and confines the target cells to a sub-region of the sensing region that at least partially overlaps the recording electrode; and using the recording electrode to record electrical activities of the confined cells. ### Claim 19 (depends on 18) The method of claim 18, wherein moving and confining comprises moving and confining the target cells to a sub-region that includes locations of minimal electrical field strength. ### Claim 20 (depends on 18) The method of claim 18, wherein moving and confining comprises moving and confining the target cells substantially near a center of the sensing region. ### Claim 21 (depends on 18) The method of claim 18, further comprising arranging at least four positioning electrodes to define a sensing region. ### Claim 22 (depends on 18) The method of claim 18, further comprising applying a pair of sinusoidal signals to generate the electric field pattern; and exposing the target cells in the sensing region to one or more dielectrophoretic forces that are generated based on the electric field pattern. ### Claim 23 (depends on 22) The method of claim 22, wherein applying a pair of sinusoidal signals includes applying a pair of alternating current signals; and exposing the target cells to one or more dielectrophoretic forces includes exposing the target cells to a negative dielectrophoretic force. ### Claim 24 (depends on 23) The method of claim 23, further comprising exposing the target cells to a negative dielectrophoretic force to enable the target cells to form an ordered network of cells. ### Claim 25 (depends on 22) The method of claim 22, further comprising selecting a pair of sinusoidal signals that are separated by a phase angle difference of 180 degrees. ### Claim 26 (depends on 18) The method of claim 18, further comprising arranging the positioning electrodes to generate electrical potentials based on a second-order polynomial that obeys Laplace's equation. ### Claim 27 (depends on 26) The method of claim 26, further comprising shaping the positioning electrodes based on equipotential boundaries of the generated electrical potentials. ### Claim 28 (depends on 18) The method of claim 18, further comprising the microelectrode sensing device is used to perform at least one analysis selected from a group including impedance spectroscopy analysis, cell poration, and electrochemical analysis of physiological changes. ### Claim 29 (depends on 18) The method of claim 18, further comprising using the microelectrode sensing device to record spontaneous action potentials or evoked action potentials from one or more excitable cells. ### Claim 30 (depends on 29) The method of claim 29, wherein recording the spontaneous action potentials or evoked potentials comprises recording from neuronal cells or heart cells. ### Claim 31 (depends on 18) The method of claim 18, further comprising applying a positive dielectrophoretic force through the recording electrode. ### Claim 32 A microelectrode sensing device, comprising: a substrate; and an array of microelectrode sensors formed on the substrate, each sensor comprising: a first layer formed over the substrate and patterned to comprise a sensing electrode which measures electrical activities of one or more target cells in a solution above the first layer; a second layer elevated above the first layer and patterned to comprise a plurality of positioning electrodes arranged to define a sensing region on top of the sensing electrode in which the solution is located, the positioning electrodes operable to generate an electric field pattern in the sensing region to move and confine the target cells above the sensing electrode; and an insulator material filled between the first and the second layers to electrically insulate the sensing electrode from the positioning electrodes, the insulator material shaped to define at least one channel between the first and the second layers to contain the solution, wherein the sensing region is located in the channel. ### Claim 33 (depends on 32) The microelectrode sensing device of claim 32, further comprising at least four positioning electrodes. ### Claim 34 (depends on 33) The microelectrode sensing device of claim 33, wherein four of the positioning electrodes are configured to form a first pair of electrodes and a second pair of electrodes operable to apply a first signal and a second signal to generate the electric field pattern; and expose the target cells in the sensing region to one or more dielectrophoretic forces that are generated based on the electric field pattern. ### Claim 35 (depends on 34) The microelectrode sensing device of claim 34, wherein the first and second signals comprise a pair of alternating current signals; and the one or more dielectrophoretic forces generated comprises a negative dielectrophoretic force. ### Claim 36 (depends on 32) The microelectrode sensing device of claim 32, wherein the recording electrode is further configured to apply a positive dielectrophoretic force. ### Claim 37 (depends on 32) The microelectrode sensing device of claim 32, further comprising a signal generator configured to apply one or more signals through the positioning electrodes or the recording electrode. ### Claim 38 A microelectrode sensing device, comprising: a substrate; and an array of microelectrode sensors formed on the substrate, each sensor comprising a first layer, that at least partially conducts electricity, formed above the substrate and patterned to comprise a recording electrode to measure electrical activities of one or more target cells in a solution; a second layer elevated above the first layer and patterned to comprise a plurality of positioning electrodes arranged to define a sensing region above the recording electrode in which the solution is located, the positioning electrodes operable to generate an electric field pattern in the sensing region to move and confine the target cells to a sub-region of the sensing region that at least partially overlaps the recording electrode; and an insulator material to electrically insulate the recording electrode from the positioning electrodes. ### Claim 39 (depends on 38) The microelectrode sensing device of claim 38, further comprising at least four positioning electrodes. ### Claim 40 (depends on 39) The microelectrode sensing device of claim 39, wherein four of the positioning electrodes are configured to form a first pair of electrodes and a second pair of electrodes operable to apply a first signal and a second signal to generate the electric field pattern; and expose the target cells in the sensing region to one or more dielectrophoretic forces that are generated based on the electric field pattern. ### Claim 41 (depends on 40) The microelectrode sensing device of claim 40, wherein the first and second signals comprise a pair of alternating current signals; and the one or more dielectrophoretic forces generated comprises a negative dielectrophoretic force. ### Claim 42 (depends on 38) The microelectrode sensing device of claim 38, wherein the recording electrode is further configured to apply a positive dielectrophoretic force. ### Claim 43 (depends on 38) The microelectrode sensing device of claim 38, further comprising a signal generator configured to apply one or more signals through the positioning electrodes or the recording electrode.