A mixture of DOPC and PE-Rhodamine B lipids (1 mg/ml; 99.5:0.5 mol% respectively) in chloroform was pipetted onto the conductive side of an ITO coated slide (~100 μL). This was spread using a cover slip to create a lipid film, which was further dried in a dessicator overnight. A PDMS spacer was fixed around the lipid film, which was hydrated with a PBS/Sucrose buffer (0.5 M; 0.5 ml). A second ITO slide was placed over the spacer such that its conductive side faced toward the lipid film. Two clips were attached to seal the slides, and electrodes connected to a generator placed on each. The sandwich was inverted (lipid film facing down) and heated to 65 °C. The volatage was set to 1.5 V and the amplitude set to 10 Hz for the first 2 hours, then at 2 Hz for the final hour. The resultant GUVs were decanted into an Eppendorf to await further use.