Using multi-frequency cross-correlation fluorometry, the monomer fluorescence lifetime of 1-palmitoyl-2-[10-(1-rmpyrenyl)decanoyl)phosphatidylcholine (Py-PC) was employed to determine the lateral diffusion constant (DT) of dioleoylphosphatidylethanolamine (DOPE) in both the lamellar (Lα) and the inverted hexagonal (HII) phases. The values of DT increased with temperature in both phases. However, the rate of increase of DT declined abruptly at ∼ 10–13°C (Lα-HII) transition temperature), as indicated by the existence of an inflection point in the log (Full-size image (<1 K)) vs. Full-size image (<1 K) plot. This observation suggests that the translational motion of lipids in the HII phase is lower than that in the Lα phase upon temperature extrapolation. Lipid perturbants, cholesterol and diacylglycerol, were found to destabilize the Lα phase of DOPE. This was demonstrated by a down-shift of the inflection point in the log (Full-size image (<1 K)) vs. Full-size image (<1 K) plot in the presence of the perturbants. Both cholesterol and 1,2-dioleoyl-sn-glycerol (diolein) decreased the lateral diffusion constant in both phases. Diolein promoted the HII phase more effectively than did the cholesterol. This is explained by an intrinsic wedge-shape geometry of diolein which strongly favors the formation of inverted cylindrical packing of the lipids.