Lutein was loaded into liposomes, and their stability against environmental stress was investigated. Subsequently, these findings were correlated with the interactions between lutein and lipid bilayer. Results showed that the liposomes with loaded lutein at concentrations of 1 and 2% remained stable during preparation, heating, storage, and surfactant dissolution. However, with further increase in the loading concentration to 5 and 10%, the stabilization role of lutein on membrane was not pronounced or even opposite. Membrane fluidity demonstrated that at 1 and 2%, lutein displayed less fluidizing properties both in the headgroup region and in the hydrophobic core of the liposome, whereas this effect was not significant at 5 and 10%. Raman spectra demonstrated that lutein incorporation greatly affected the lateral packing order between acyl chains and longitudinal packing order of lipid acyl chains. These results may guide the potential application of liposomes as carriers for lutein in nutraceuticals and functional foods.