We have used steady state and time-resolved fluorescence spectroscopy in concert with TEM to study organization and dynamics of molecules comprising liposomes, discoidal micelles, and spherical micelles. The lipid aggregates contained controlled amounts of lipids with headgroups modified with a thiol-terminated polyethylene glycol (thio-PEG lipids) and a small amount of 1-palmitoyl-2-(pyrene-1-yl)decanoyl-sn-glycero-3-phosphocholine (pyrene tethered DPPC), pyrene, or perylene as spectroscopic probes. The maximum diameter of the lipid aggregates was controlled by the polycarbonate Filter pore size used in the extrusion process. The concentration of thio-PEG lipid in the aggregates determines not only the shape of the lipid assemblies but also the organization of the molecules within the assembly. Fluorescence lifetime and anisotropy decay data show that the immediate environment of pyrene tethered DPPC changes with the addition of thio-PEG lipid. In contrast, the dynamics of free chromophore (perylene) are insensitive to the addition of thio-PEG lipid. The addition of thio-PEG lipid to the lipid assembly produces changes in organization that are most pronounced in the lipid headgroup region. Reorientation dynamics of perylene show that the organization of the lipid bilayer acyl chain region is affected little by the addition of thio-PEG lipid and consequent macroscopic changes in the morphology of the lipid assemblies.