Endothelial cell electrical impedance parameter artifacts produced by a gold electrode and phase sensitive detection.

Frequency dependent cellular micro-impedance estimates obtained from a gold two-electrode configuration using phase sensitive detection have become increasingly used to evaluate cellular barrier model parameters. The results of this study show that cellular barrier function parameter estimates optimized using measurements obtained from this biosensor are highly susceptible to both time dependent and systematic instrumental artifacts. Based on a power spectral analysis of experimentally measured microelectrode voltages, synchronous, 60 Hz, and white Gaussian noise were identified as the most significant time dependent instrumental artifacts. The reduction of these artifacts using digital filtering produced a corresponding reduction in the optimized model parameter fluctuations. Using a series of instrumental circuit models, this study also shows that electrode impedance voltage divider effects and circuit capacitances can produce systematic deviations in cellular barrier function parameter estimates. Although the implementation of an active current source reduced the voltage divider effects, artifacts produced by coaxial cable and other circuit capacitive elements at frequencies exceeding 1 kHz still remained. Reducing time dependent instrumental fluctuations and systematic errors produced a significant reduction in cellular model barrier parameter errors and improved the model fit to experimental data.

Modeling error and stability of endothelial cytoskeletal membrane parameters based on modeling transendothelial impedance as resistor and capacitor in series.

Transendothelial impedance across an endothelial monolayer grown on a microelectrode has previously been modeled as a repeating pattern of disks in which the electrical circuit consists of a resistor and capacitor in series. Although this numerical model breaks down barrier function into measurements of cell-cell adhesion, cell-matrix adhesion, and membrane capacitance, such solution parameters can be inaccurate without understanding model stability and error. In this study, we have evaluated modeling stability and error by using a chi(2) evaluation and Levenberg-Marquardt nonlinear least-squares (LM-NLS) method of the real and/or imaginary data in which the experimental measurement is compared with the calculated measurement derived by the model. Modeling stability and error were dependent on current frequency and the type of experimental data modeled. Solution parameters of cell-matrix adhesion were most susceptible to modeling instability. Furthermore, the LM-NLS method displayed frequency-dependent instability of the solution parameters, regardless of whether the real or imaginary data were analyzed. However, the LM-NLS method identified stable and reproducible solution parameters between all types of experimental data when a defined frequency spectrum of the entire data set was selected on the basis of a criterion of minimizing error. The frequency bandwidth that produced stable solution parameters varied greatly among different data types. Thus a numerical model based on characterizing transendothelial impedance as a resistor and capacitor in series and as a repeating pattern of disks is not sufficient to characterize the entire frequency spectrum of experimental transendothelial impedance.