Free-standing microcrystals of an organic fluorescent dye, specifically, 4-n-octylamino-7-nitrobenz-2-oxa-1,3-diazole, were prepared using a solvent-exchange process at room temperature, in the presence of polymers used as additives. Parameters such as the dye concentration, the nature and concentration of the polymer, and the pH of the solution were varied. Six samples of microcrystals were therefore obtained and characterized by fluorescence microscopy and by electron microscopy (TEM and SEM). They differed by their content in microcrystals, the shape and size of which depends strongly on experimental conditions. Curiously, the UV/vis absorption spectra of the microcrystal suspensions were very different from one sample to another. As a result the emission spectra were also varied. The diversity of the optical response obtained was attributed to the presence of several dye populations in the microcrystal suspensions. A distinction was made between the intrinsic spectral properties of the microcrystals and artifacts due to the presence of the additives.