Advancing membrane biology: single-molecule approaches meet model membrane systems

Model membrane systems have emerged as essential platforms for investigating membrane-associated processes in controlled environments, mimicking biological membranes without the complexity of cellular systems. However, integrating these model systems with single-molecule techniques remains challenging due to the fluidity of lipid membranes, including undulations and the lateral mobility of lipids and proteins. This mini-review explores the evolution of various model membranes ranging from black lipid membranes to nanodiscs and giant unilamellar vesicles as they adapt to accommodate electrophysiology, force spectroscopy, and fluorescence microscopy. We highlight recent advancements, including innovations in force spectroscopy and single-molecule imaging using free-standing lipid bilayers, and the development of membrane platforms with tunable composition and curvature for improving fluorescence-based studies of protein dynamics. These integrated approaches have provided deep insights into ion channel function, membrane fusion, protein mechanics, and protein dynamics. We highlight how the synergy between single-molecule techniques and model membranes enhances our understanding of complex cellular processes, paving the way for future discoveries in membrane biology and biophysics. [BMB Reports 2025; 58(1): 33-40].