Label-free monitoring of excitation-contraction coupling and excitable cells using impedance based systems with millisecond time resolution

Abstract

A method for monitoring a cell beating parameter, which includes adding excitable cells capable of beating to a system for monitoring cell-substrate impedance, the system having an electrode array electrically coupled to an impedance analyzer that measures cell-substrate impedance at millisecond time resolution, and a software program that determines a cell beating parameter from measured cell-substrate impedance; monitoring cell-substrate impedance of the excitable cells; and determining the cell beating parameter from the monitored cell-substrate impedance.

What is claimed is: 
     
         1 . A method for monitoring a cell beating parameter, the method comprising:
 a) adding excitable cells capable of beating to a system for monitoring cell-substrate impedance, the system comprising an electrode array electrically coupled to an impedance analyzer that measures cell-substrate impedance at millisecond time resolution, and a software program that determines a cell beating parameter from measured cell-substrate impedance;   b) monitoring cell-substrate impedance of the excitable cells; and   c) determining the cell beating parameter from the monitored cell-substrate impedance.   
     
     
         2 . The method of  claim 1 , wherein the cells are cardiomyocytes or stem cells. 
     
     
         3 . The method of  claim 1 , wherein the cells comprise a gene knockout, a gene knockdown or express a transgene. 
     
     
         4 . The method of  claim 1 , wherein the cell-substrate impedance is resolved between two consecutive time points that are within 20 milliseconds apart. 
     
     
         5 . The method of  claim 1 , wherein the cell-substrate impedance is resolved between two consecutive time points that are within 10 milliseconds apart. 
     
     
         7 . The method of  claim 1  further comprising adding an agent suspected of affecting cell beating to the cells, wherein cell-substrate impedance is monitored at millisecond time resolution before and after adding the agent. 
     
     
         8 . The method of  claim 7 , further comprising determining the beating parameter from the monitored cell-substrate impedance before and after adding the agent and comparing the beating parameters to assess whether the agent affected cell beating. 
     
     
         9 . The method of  claim 1 , wherein the beating parameter is selected from the group consisting of a beating cycle, a beating amplitude and a beating rate. 
     
     
         10 . The method of  claim 1 , wherein the beating parameter is a rise or a decay of a beating cycle. 
     
     
         11 . The method of  claim 1 , wherein the beating parameter is a beating cycle peak. 
     
     
         12 . The method of  claim 1 , wherein the beating parameter is determined from an impedance-based curve. 
     
     
         13 . The method of  claim 1 , wherein the cells are added to at least two wells that each have a separate electrode array, the method further comprising adding an agent suspected of affecting cell beating to one of the wells to form a test well and maintaining another well as a control well without the agent, and wherein the steps of monitoring cell-substrate impedance and determining the beating parameter is performed for both wells, and the method further comprising comparing the beating parameters to assess whether the agent affected the cell beating parameter. 
     
     
         14 . The method of  claim 13 , wherein the beating parameter is selected from the group consisting of a beating cycle, a beating amplitude and a beating rate. 
     
     
         15 . The method of  claim 13 , wherein the beating parameter is a rise or a decay of a beating cycle. 
     
     
         16 . The method of  claim 13 , wherein the beating parameter is a beating cycle peak. 
     
     
         17 . The method of  claim 13 , wherein the beating parameter is determined from an impedance-based curve.