The UV/Vis absorption band maximum lambda(max) of trans-4,4-nitrophenylaminoizobenzene, the thermal isomerization rate constant k(iso) of its cis-isomer, the fluorescence intensity ratio of monomer and excimer, and the fluorescence lifetime of the excimer, respectively, of 1,3-di(1-pyrenyl)propane were determined as probes for polarity, water content, and viscosity, respectively, in unilamellar vesicles of din-alkyl-dimethylammonium bromides and 1,2-acyl-sn-glycero-3-phosphocholines. The dependence on vesicle size, the solvent (water or HEPES buffer/NaCl solution, each with H2O or D2O), and the temperature (20-60 degreesC) was studied. Apparent Arrhenius activation energies and kinetic solvent isotope effects (KSIE = k(iso), (H2O)/k(iso), (D2O)) were derived. Size and stability of the vesicles prepared by extrusion were controlled by dynamic light scattering. The probe properties clearly indicate the reversibly decreasing size of didodecyldimethylammonium bromide vesicles with increasing temperature but are insensitive against vesicles size variation in most other cases. In the temperature range of the main phase transition of the bilayers, changes of the microenvironment of the probes, and their changing position in the bilayer, respectively, are reflected by characteristic changes of their properties. Buffer/NaCl solution causes vanishing influence of the lipid chain but remaining difference between cationic and zwitterionic headgroups probed by means of k(iso). (C) 2004 Elsevier Inc. All rights reserved.