Sensor spacing affects the tissue impedance spectra of rabbit ventricular epicardium.

This study was designed to test the hypothesis that a complex composite impedance spectra develops when stimulation and recording of cardiac muscle with sufficiently fine spatial resolution in a four-electrode configuration is used. With traditional (millimeter scale) separations, the ratio between the recorded interstitial central potential difference and total supplied interstitial current is constant at all frequencies. This occurs because the fraction of supplied current that redistributes to the intracellular compartment depends on effective membrane resistance between electrodes, which is low, to a much greater extent than effective membrane capacitance. The spectra should therefore change with finer separations at which effective membrane resistance increases, as supplied current will remain primarily interstitial at lower frequencies and redistribute between compartments at higher frequencies. To test this hypothesis, we built arrays with sensors separated (d) by 804 µm, 452 µm, and 252 µm; positioned those arrays across myocyte axes on rabbit ventricular epicardium; and resolved spectra in terms of resistivity (ρt) and reactivity (χt) over the 10 Hz to 4,000 Hz range. With all separations, we measured comparable spectra with predictions from passive membrane simulations that used a three-dimensional structural framework in which intracellular, interstitial, and membrane properties were prescribed based on the limited data available from the literature. At the finest separation, we found mean ρt at 100 Hz and 4,000 Hz that lowered from 395 Ω-cm to 236 Ω-cm, respectively, with maximal mean χt of 160 Ω-cm. This experimental confirmation of spectra development in whole heart experiments is important because such development is central to achieve measurements of intracellular and interstitial passive electrical properties in cardiac electrophysiological experiments using only interstitial access.

A new approach for resolution of complex tissue impedance spectra in hearts.

This study was designed to test the feasibility of using sinusoidal approximation in combination with a new instrumentation approach to resolve complex impedance (uCI) spectra from heart preparations. To assess that feasibility, we applied stimuli in the 10-4000 Hz range and recorded potential differences (uPDs) in a four-electrode configuration that allowed identification of probe constants (Kp) during calibration that were in turn used to measure total tissue resistivity ρt from rabbit ventricular epicardium. Simultaneous acquisition of a signal proportional to the supplied current (Vstim) with uPD allowed identification of the V- I ratio needed for ρt measurement, as well as the phase shift from Vstim to uPD needed for uCI spectra resolution. Performance with components integrated to reduce noise in cardiac electrophysiologic experiments, in particular, and provide accurate electrometer-based measurements, in general, was first characterized in tests using passive loads. Load tests showed accurate uCI recovery with mean uPD SNRs between 10 (1) and 10 (3) measured with supplied currents as low as 10 nA. Comparable performance characteristics were identified during calibration of nine arrays built with 250 µm Ag/AgCl electrodes, with uCIs that matched analytic predictions and no apparent effect of frequency ( F = 0.12, P = 0.99). The potential ability of parasitic capacitance in the presence of the electrode-electrolyte interface associated with the small sensors to influence the uCI spectra was therefore limited by the instrumentation. Resolution of uCI spectra in rabbit ventricle allowed measurement of ρt = 134 ± 53 Ω· cm. The rapid identification available with this strategy provides an opportunity for new interpretations of the uCI spectra to improve quantification of disease-, region-, tissue-, and species-dependent intercellular uncoupling in hearts.