The tubular system of cardiomyocytes plays a key role in excitation-contraction coupling. To determine the area of the tubular membrane in relation to the area of the surface membrane, indirect measurements through the determination of membrane capacitances by electrophysiological measurements are currently used in addition to microscopic methods. Unlike existing electrophysiological methods based on an irreversible procedure (osmotic shock), the proposed approach uses a reversible short-term intermittent increase in the electrical resistance of the extracellular medium. The resulting increase in the lumen resistance of the tubular system makes it possible to determine separately capacitances of the tubular and surface membranes from altered capacitive current responses to subthreshold voltage-clamped rectangular pulses. Based on the analysis of the time course of capacitive current, computational relations were derived which allow to quantify elements of the electrical equivalent circuit of the measured cardiomyocyte including both capacitances. The exposition to isotonic low-conductivity sucrose solution is reversible which is the main advantage of the proposed approach allowing repetitive measurements on the same cell under control and sucrose solutions. In addition, it might be possible to identify changes in both surface and tubular membrane capacitances caused by various interventions. Preliminary experiments in rat ventricular cardiomyocytes (n = 10) resulted in values of the surface and tubular capacitances 72.3 ± 16.4 and 42.1 ± 14.7 pF, respectively, implying the fraction of tubular capacitance/area of 0.36 ± 0.08. We conclude that the newly proposed method provides results comparable to those reported in literature and, in contrast to the currently used methods, enables repetitive evaluation of parameters describing the surface and tubular membranes. It may be used to study alterations of the tubular system resulting from various interventions including associated cardiac pathologies.