We investigated the effects of the alignment and ordering of pi-conjugated perfluorinated dendrimers containing pyrene moieties in their cores on their photoluminescence (PL) properties. The pyrene molecules are stacked in columns surrounded by aromatic and semifluorinated tails, which can be conjugated and act as chromophores. Polarized light microscopy (PLM), cross-sectional scanning electron microscopy (SEM), and atomic force microscopy (AFM) results show that variation of the cooling rate of the dendrimers produces variation in their orientation and ordering: Slow cooling (similar to < 0.5 degrees C/min) of the isotropic melt in a sandwich glass cell results in a high degree of ordering and the vertical alignment of the columns on the substrate, in which the stacked pyrene molecules are oriented parallel to the surface over large areas. In contrast, rapid cooling (similar to > 10 degrees C/min) leads to the planar alignment of the columns with significant disorder on the same substrate. UV-vis, PL, SEM, and AFM results show that the quenched columns with a planar orientation produce a broad emission band and a second weak shoulder, which indicates the presence of isolated molecules. However, the high degree of ordering of the columns with a vertical alignment produces a red-shift in the PL spectrum, with very few isolated molecules. By comparing two films with different alignments but similar ordering, we show that the ordering of this material has a greater influence on the PL spectrum than the alignment. This effect of the ordering of the columns was further verified by comparing the optical properties of the isolated dendrimers with those of small pi-conjugated molecules in solution and solid films.