Real time monitoring of endothelial cell actin filament disruption by cytochalasin D using a cellular impedance biosensor

Using a novel cellular impedance biosensor and confocal microscopy, this study has examined the dynamic and steady state cellular impedance response of porcine pulmonary endothelial cells to varying doses of cytochalasin D. Endothelial cell monolayer impedances were obtained using an array of gold microelectrodes coated with fibronectin to facilitate endothelial cell adhesion. Impedance measurements were acquired at 5.6 kHz by phase sensitive detection from a lock-in amplifier. The electrically measured cytochalasin D dose dependant actin disruption was successfully correlated with actin stained confocal microscopy images quantified using image-processing techniques. Based on this study, the cellular kinetic response to cytochalasin D increased systematically with the dose and saturated at a critical concentration. The real time quantification of pharmacological agents that target specific elements of the cytoskeleton using electrical impedance measurements therefore has a number of important applications in understanding the dynamic and steady state response of endothelial cells to toxins and drug induced cellular cytoskeletal micromechanics