We achieved single-molecule surface-enhanced Raman scattering (SM-SERS) spectra from ultralow concentrations (10−15 M) of fullerene C60 on uniformly assembled Au nanoparticles. It was found that resonant excitation at 785 nm is a powerful tool to probe SM-SERS in this system. The appearance of additional bands and splitting of some vibrational modes were observed because of the symmetry reduction of the adsorbed molecule and a relaxation in the surface selection rules. Time-evolved spectral fluctuation and ‘hot spot’ dependence in the SM-SERS spectra were demonstrated to result from the single-molecule Raman behavior of the spherical C60 on Au nanoparticles. Copyright © 2010 John Wiley & Sons, Ltd.