Palmitic acid and cholesterol have been shown to form, under certain conditions, bilayers in the liquid ordered (lo) phase. In the present work, the contribution of the hydrophobic match between cholesterol (chol), and the acyl chain of saturated fatty acids (FA) has been examined. The behavior of FA/chol mixtures where the FA acyl chain length was varied between 12 and 24 carbon atoms was investigated by infrared and H-2 NMR spectroscopy, as well as by differential scanning calorimetry. It was found that only fatty acids with acyl chains of 14-18 carbon atoms lead to the formation of lo phase bilayers. The length of these chains corresponds, in fact, to the length of the long axis of the cholesterol molecule. Therefore, the hydrophobic match between the apolar parts of the molecular constituents appears to be a requisite for the formation of lamellar lo phases.