Simultaneous impedance spectroscopy and fluorescence microscopy for the real-time monitoring of the response of cells to drugs.

A dual fluorescence microscopy and electrochemical strategy to investigate how cell-surface interactions influence the cellular responses to cues for the cell-based biosensing of drug efficacy is reported herein. The combined method can be used to not only monitor the importance of controlling the cellular adhesive environment on the cell response to drugs but it also provides biological information on the timescales of downstream outside-in signaling from soluble cues. As an example of the use of the combined method, we show how adhesive cues influence the signalling responses of cells to soluble cues. G-protein-coupled receptors were used as the target for the soluble cues. The changes in cell adhesion, cell morphology and Ca flux induced by soluble histamine were simultaneously monitored as a function of the spacing of the adhesive ligand RGD on the interdigitated indium tin oxide electrodes. The simultaneous measurements revealed that the timescales of histamine-induced Ca mobilization and the decrease in cell-cell adhesions are correlated. Furthermore, cells on the surfaces with an RGD spacing of 31 nm were shown to display a faster release of Ca and change in cell adhesion upon histamine stimulation compared to cells on other surfaces.