Abstract: Devices and methods for performing extracellular recording of cells, such as excitable cells, cardiomyocytes, and cardiomyocyte precursor cells is provided. An exemplary device includes a nonconductive substrate forming or provided as a base of one or more wells; a recording electrode positioned on the substrate within the well, wherein the recording electrode is accessible to cells when a cell sample is added to the device; and a reference electrode positioned within the well in a cell-free zone, the cell-free zone characterized as free from contact with cells when the cell sample is added to the device, thereby preventing contact between cells and the reference electrode.

Abstract: Devices, systems and methods for monitoring excitable cells, such as cardiomyocytes, on microelectrode arrays that couple the electro-stimulation of excitable cells to induce or regulate cardiomyocyte beating and the simultaneous measurement of impedance and extracellular recording to assess changes in cardiomyocyte beating, viability, morphology or electrophysical properties in response to a plurality of treatments.

Abstract: A system for monitoring cell-substrate impedance of excitable cells and monitoring the beating cycle of cardiomyocytes, the system including: a device for monitoring cell-substrate impedance at 20 millisecond time resolution, the device having one or more wells on a nonconductive substrate, wherein at least one well of the one or more wells comprises an electrode array fabricated on the substrate for measurement of cell-substrate impedance at 20 millisecond time resolution; an impedance analyzer that measures cell-substrate impedance from the at least one well at 20 millisecond time resolution; electronic circuitry that electrically connects the electrode array from the at least one well to the impedance analyzer; and a software program that analyzes the measured cell-substrate impedance.

Abstract: A method of identifying a therapeutic compound for treating cancer in a human subject, the method including: providing a device that measures cell-substrate impedance; culturing cancer cells in the at least two wells, wherein the cancer cells are obtained from a human subject and have a receptor tyrosine kinase (RTK) pathway; adding to a first well a proposed therapeutic compound that affects an RTK pathway and an RTK stimulating factor for the RTK pathway to form a test well, and adding to another well the RTK stimulating factor to form a control well; continuously monitoring cell-substrate impedance of the at least two wells; and determining a difference in impedance or optionally in cell index between the test well and control well; and if significantly different, concluding the proposed therapeutic compound is therapeutically active in the RTK pathway within the cancer cells of the human subject.

Abstract: Use of impedance devices in methods of generating a time dependent cellular profiles (TCRP) for the modulation of oncogene addicted cells and comparing the impedance-based TCRP to controls or knowns to identify signature time dependent cellular profiles.

Abstract: A method of identifying a potential therapeutic compound that affects a Receptor Tyrosine Kinase (RTK) pathway in cancer cells, which includes: providing a device capable of measuring cell-substrate impedance; culturing cancer cells in serum-free media in at least two wells of the device; adding to a first well a proposed therapeutic compound that affects a RTK pathway and a RTK stimulating factor for the RTK pathway to form a test well, and adding to another well the RTK stimulating factor to form a control well; continuously monitoring cell-substrate impedance of the at least two wells and optionally determining cell indices from the monitored cell-substrate impedance; and determining a difference in impedance or optionally cell index between the test well and control well; and if significantly different, concluding the proposed therapeutic compound is therapeutically active in the RTK pathway within the cancer cells.

Abstract: A system for monitoring cell-substrate impedance of excitable cells and monitoring the beating cycle of cardiomyocytes, the system including: a device for monitoring cell-substrate impedance at 20 millisecond time resolution, the device having one or more wells on a nonconductive substrate, wherein at least one well of the one or more wells comprises an electrode array fabricated on the substrate for measurement of cell-substrate impedance at 20 millisecond time resolution; an impedance analyzer that measures cell-substrate impedance from the at least one well at 20 millisecond time resolution; electronic circuitry that electrically connects the electrode array from the at least one well to the impedance analyzer; and a software program that analyzes the measured cell-substrate impedance.

Abstract: Use of cell-substrate impedance based methods for screening for agonists of G-Protein Coupled Receptors (GPCRs) or inhibitors of a Receptor Tyrosine Kinases (RTKs), identifying compounds that affect GPCR or RTK pathways, validating molecular targets involved in a GPCR or RTK signaling pathways, monitoring dose-dependent functional activation of GPCR or RTK; determining desensitization of a GPCR and identifying a compound capable of affecting RTK activity in cancer cell proliferation.

Abstract: Methods of assessing cytolysis of cancer cells, including providing a cell-substrate impedance monitoring device operably connected to an impedance analyzer, wherein the device comprises a well for receiving cells and an electrode array at a base of the well; adding target cells characterized as cancer cells to the well; adding effector cells to the well to form a test well, wherein the effector cells are immune cells obtained or derived from a same patient as the target cells; monitoring cell-substrate impedance of the test well before and after adding the effector cells and optionally deriving an impedance-based parameter from the impedance; and determining effectiveness of effector cell killing of the target cells by comparing the impedance or impedance based parameter over time.

Abstract: A system for monitoring impedance of excitable cells in vitro, which includes a device for monitoring cell-substrate impedance at 20 millisecond resolution, which includes a nonconductive substrate with one or more electrode arrays fabricated in one or more wells, wherein cell attachment on the substrate can result in a detectable change in impedance between electrodes within each electrode array; an impedance analyzer capable of impedance measurement at 20 millisecond time resolution; electronic circuitry that can engage said device and selectively connect said two or more electrode arrays of said device to said impedance analyzer; and a software program that controls said electronic circuitry and records and analyzes data obtained from said impedance analyzer.

Abstract: A system for monitoring impedance of excitable cells in vitro, which includes a device for monitoring cell-substrate impedance at 20 millisecond resolution, which includes a nonconductive substrate with one or more electrode arrays fabricated in one or more wells, wherein cell attachment on the substrate can result in a detectable change in impedance between electrodes within each electrode array; an impedance analyzer capable of impedance measurement at 20 millisecond time resolution; electronic circuitry that can engage said device and selectively connect said two or more electrode arrays of said device to said impedance analyzer; and a software program that controls said electronic circuitry and records and analyzes data obtained from said impedance analyzer.

Abstract: A method of identifying a potential therapeutic compound that affects a Receptor Tyrosine Kinase (RTK) pathway in cancer cells, which includes: providing a device capable of measuring cell-substrate impedance; culturing cancer cells in serum-free media in at least two wells of the device; adding to a first well a proposed therapeutic compound that affects a RTK pathway and a RTK stimulating factor for the RTK pathway to form a test well, and adding to another well the RTK stimulating factor to form a control well; continuously monitoring cell-substrate impedance of the at least two wells and optionally determining cell indices from the monitored cell-substrate impedance; and determining a difference in impedance or optionally cell index between the test well and control well; and if significantly different, concluding the proposed therapeutic compound is therapeutically active in the RTK pathway within the cancer cells.

Abstract: Methods for identifying compounds that modulate cellular responses stimulated by IgE, which include providing an impedance-based system that monitors cell-substrate impedance of cells on a substrate; introducing cells to the substrate of the system; adding at least one test compound and IgE to the cells, wherein the at least one test compound is suspected of modulating cell responses stimulated by the IgE; adding an antigen to the cells; monitoring the cell-substrate impedance of cells on the substrate; and analyzing the cell-substrate impedance to evaluate whether the at least one test compound alters a cellular response to stimulation with the IgE.

Abstract: Methods of identifying whether a biologically active agent affects an oncogene addicted pathway within a cancer cell, by introducing a biologically active agent suspected of affecting an oncogene addicted pathway to a first well and a negative control to a second well, and introducing a stimulating agent that stimulates the oncogene addicted pathway to both wells; monitoring cell-substrate impedance of the two wells and optionally determining cell indices from impedance values; generating an impedance based curve for each of the two wells from the impedance values or from the cell indices; comparing the impedance-based curves to determine a degree of similarity; and if significantly different concluding the biologically active agent affects the oncogene addicted pathway within the cancer cells.

Abstract: A method of determining a beating parameter of cells that undergo excitation contraction coupling, the method including providing a cell analysis device having a substrate and a sensor that measures cell adhesion or attachment to the substrate in millisecond time resolution; adding excitable cells capable of undergoing excitation contraction coupling to the substrate; monitoring cell adhesion or attachment of the excitable cells to the substrate in millisecond time resolution; and calculating one or more beating parameters from the monitored adhesion.

Abstract: Systems and methods for improved monitoring of excitation-contraction coupling and excitable cells are provided, which provide millisecond time resolution. The system is capable of continuously monitoring excitation-contraction coupling in a relatively high-throughput manner. The system includes a device for monitoring cell-substrate impedance, an impedance analyzer capable of impedance measurements at millisecond time resolution, electronic circuitry that can engage the device and selectively connect two or more electrode arrays of the device to the impedance analyzer and a software program that controls the electronic circuitry and records and analyzes data obtained from the impedance analyzer.

Abstract: The present invention includes devices, systems, and methods for assaying cells using cell-substrate impedance monitoring. In one aspect, the invention provides cell-substrate impedance monitoring devices that comprise electrode arrays on a nonconducting substrate, in which each of the arrays has an approximately uniform electrode resistance across the entire array. In another aspect, the invention provides cellular assays that use impedance monitoring to detect changes in cell behavior or state. In some preferred aspects, the assays are designed to investigate the affects of compounds on cells, such as cytotoxicity assays.

Abstract: A method of determining one or more beating parameters for use in cardiomyocyte beating analysis including: providing a cell analysis device including wells, each well including a sensor capable of monitoring beating of cardiomyoctes in millisecond time resolution; adding cardiomyocytes to the wells; monitoring the beating of the cardiomyocytes in millisecond time resolution to obtain a plurality of beating measurements; and calculating one or more beating parameters from the plurality of beating measurements.

Abstract: A method of performing an assay of a response of two or more cell types to a test compound, including: providing a device for monitoring cell-substrate impedance, adding at least two different cell types to the device; adding at least one test compound to form at least two test compound wells; providing at least two control wells; monitoring impedance of the at least two test compound wells and of the at least two control wells at three or more time points after adding the at least one test compound; and analyzing measured impedance from the at least two test compound wells and from the at least two control wells at the three or more time points to obtain information on the response of the different cell types to the at least one test compound.

Abstract: An insert device and co-culture device assembly incorporating the insert device, which includes an insert chamber with two fluid impermeable side walls that extend from a microporous bottom to an open top to form an inner cavity. The first side wall is shaped to form a convex arc that follows between about half of a circumference of the well and less than an entire circumference of the well. The second side wall joins ends of the first side wall and protrudes inward towards a center of the convex arc. A flange extends outward from or beneath the top and is notched to form a gap adjacent to the second side wall, which forms an access port allowing access to the lower chamber with a micropipette tip when the insert device is inserted into a well of a single or multi-well plate.