The potential-controlled incorporation of DOPC liposomes (100 nm diameter) into an adsorbed octadecanol layer on Au(111) was studied using electrochemical and in situ fluorescence microscopy. The adsorbed layer of octadecanol included a small amount of a lipophilic fluorophore-octadecanol modified with BODIPY-to enable fluorescence imaging. The deposited octadecanol layer was found not to allow liposomes to interact unless the potential was less than -0.4 V/SCE, which introduces defects into the adsorbed layer. Small increases in the capacitance of the adsorbed layer were measured after introducing the defects, allowing the liposomes to interact with the defects and then annealing the defects at 0 V/SCE. A change in the adsorbed layer was also signified by a more positive desorption potential for the liposome-modified adsorbed layer as compared to that for an adsorbed layer that was porated in a similar fashion but without liposomes present in the electrolyte. These subtle changes in capacitance are difficult to interpret, so an in situ spectroscopic study was performed to provide a more direct measure of the interaction. The incorporation of liposomes should result in an increase in the fluorescence measured because the fluorophore should become further separated from the gold surface, reducing the efficiency of fluorescence quenching. No significant increase in the fluorescence of the adsorbed layer was observed during the potential pulses used in the poration procedure in the absence of liposomes. In the presence of liposomes, the fluorescence intensity was found to depend on the potential and time used for poration. At 0 V/SCE, no significant change in the fluorescence was observed for defect-free adsorbed layers. Changing the poration potential to -0.4 V/SCE caused significant increases in the fluorescence and the appearance of new structural features in the adsorbed layers that were more easily observed during the desorption procedure. The extent of fluorescence changes was found to be strongly dependent on the nature of the adsorbed layer under investigation, which suggests that the poration and liposome interaction are dependent on the quality of the adsorbed layer and its ease of poration through changes in the electrode potential.