Microcavity supported lipid bilayers, MSLBs, were applied to an electrochemical investigation of ionophore mediated ion transport. The arrays comprise of a 1 cm(2) gold electrode imprinted with an ordered array of uniform spherical-cap pores of 2.8 mu m diameter prepared by gold electrodeposition through polystyrene templating spheres. The pores were pre-filled with aqueous buffer prior to Langmuir-Blodgett assembly of a 1,2-dioleoylsn-glycero-3-phosphocholine bilayer. Fluorescence lifetime correlation spectroscopy enabled by the micron dimensions of the pores permitted study of lipid diffusion across single apertures, yielding a diffusion coefficient of 12.58 +/- 1.28 mu m(2) s(-1) and anomalous exponent of 1.03 +/- 0.02, consistent with Brownian motion. From FLCS, the MSLBs were stable over 3 days and electrochemical impedance spectroscopy of the membrane with and without ionic gradient over experimental windows of 6 h showed excellent stability. Two ionophores were studied at the MSLBs; Valinomycin, a K+ uniporter and Nigericin, a K+/H+ antiporter. lonophore reconstituted into the DOPC bilayer resulted in a decrease and increase in membrane resistance and capacitance respectively. Significant increases in Valinomycin and Nigericin activity were observed, reflected in large decreases in membrane resistance when K+ was present in the contacting buffer and in the presence of H+ ionic gradient across the membrane respectively. (C) 2016 Elsevier B.V. All rights reserved.