Capacitance measurements. An analysis of the phase detector technique used to study exocytosis and endocytosis

Abstract

We have studied the admittance of patch-clamped mast cells during exocytosis and found that they are adequately described by a four parameter equivalent circuit. On the basis of these measurements, we show that, contrary to current belief, when using a phase sensitive detector, small capacitance changes due to exocytosis or endocytosis should be studied by measuring current 90 degrees out of phase, relative to the component that corresponds to changes in series resistance. We have extended the theory on phase-detectors to include the errors in the estimation of step changes of membrane capacitance. We show that the measured capacitance of a secretory granule can be up to 80% smaller than its true value, during the course of a typical mast cell degranulation. We also describe a software-based phase-detector that simplifies capacitance measurements.