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Optical Multisite Detection Of Membrane Potentials In Single Cardiomyocytes During Voltage Clamp

We have performed voltage clamp experiments in single cardiomyocytes combined with simultaneous optical multisite monitoring of membrane potentials. The fluorescent potential sensitive dye di-4-ANEPPS was used for staining and an argon ion laser for excitation of fluorescence. The percentage change of the fluorescence light intensity was about 5% per lOOmV, and showed a linear relationship (within the limits of noise). Optical signals obtained simultaneously from different measuring spots on a cell sometimes showed timelags of about 100 ps. Normalizing the optical signals to the clamp steps allowed to obtain calibrated membrane potential values within a cell. This promises to be a powerful method to interpret voltage clamp experiments with large and fast currents.