Accuracy considerations for capacitance estimation by voltage steps in cardiomyocytes

Electrophysiologists routinely use simple voltage steps to evaluate cell membrane capacitance derived from corresponding current responses. Frequently, the resting membrane voltage V_rest is employed as holding potential for the subsequent command voltage step and more or less accurate methods are utilised to analyse the transient current. Another choice as holding potential is the peak of the "quasi steady-state" current to voltage relationship, V_peak. The aim of this study is the systematic evaluation of capacitance estimation accuracy from voltage step experiments depending on the choice of holding potential and analysis method. In this paper, a simulation approach is employed to analyse the current response of a model patch-clamp circuit. Four commonly accepted methods are implemented, utilizing different aspects of the transient current (charge, membrane time constant, and influence of the series resistance) in various combinations and with various degrees of refinement. This simulation study indicates an acceptable accuracy of the elaborated methods for capacitance estimation at holding potentials V_rest and V_peak over a broad range of capacitance as well as series resistance values. Simple integration of the current transient provides sufficient accuracy at holding potentials, which effectively minimizes changes in resistive membrane current flow during command voltage steps (particularly around V_peak). However, biphasic command protocols performed at V_peak activate voltage dependent sodium channels, thereby possibly leading to the threshold voltage for an action potential. Compared to V_rest, all methods utilizing monophasic step protocols, gain additional accuracy, when applied at V_peak as holding potential.