Photophysical studies have been carried out to explore the aggregation behavior of coumarin-153 (C153) dye in polar organic solvents of both aprotic and protic nature, namely acetonitrile (ACN) and ethanol (EtOH). No unusual behavior is observed in aprotic ACN solvent, suggesting only themonomers of the dye as the single emitting species in the solution. In protic EtOH solvent, however, the dye shows the presence of multiple emitting species in the solution. The concentration-, temperature- and wavelength-dependent changes in the fluorescence decays, and the time-resolved emission spectra (TRES) and area-normalized emission spectra (TRANES) suggest the coexistence of dye aggregates along with the dye monomers in the EtOH solution. Observed results indicate that the emission spectra of the aggregates are substantially blueshifted compared to the spectra of the monomers, suggesting the H-aggregation of the dye in the present cases. Time-resolved fluorescence anisotropy, ultrafast fluorescence up-conversion measurements and scanning electron microscopy studies support the aggregation of the dye in EtOH solution. Strong dipole−dipole interaction is supposedly responsible for the aggregation of C153 dye (dipole moment ~6.4 D) and the polar protic solvent EtOH apparently stabilizes the aggregates through solute−solvent hydrogen bonding interaction, which is not possible in aprotic ACN solvent. This is further supported by the time-resolved fluorescence results in a strongly hydrogen bond donating solvent, 2,2,2-trifluoroethanol. Aggregation of C153 dye observed in the present study in polar protic organic solvent is an intriguing finding, as the dye is widely used as a fluorescent probe for various photochemical studies, where overlooking such aggregation will definitely mislead the observed results.