Correlated AFM and NanoSIMS imaging to probe cholesterol-induced changes in phase behavior and non-ideal mixing in ternary lipid membranes

►Lo-Ld phase separation does not occur in DLPC/DSPC membranes containing cholesterol. ►Correlated AFM and NanoSIMS imaging identifies lipid domain structure and composition. ►Cholesterol addition causes elongation of gel-phase domains in DLPC/DSPC membranes. ►Ordered domains in DLPC/DSPC membranes with ≤ 19 mol% cholesterol contain < 7 mol% DLPC.