We have investigated the effects of substrate topography on the nucleation and growth behavior of 321 stainless steel (ss) films, introducing textured surfaces into mica substrates by the abrasion treatment. In this study, two groups of samples were prepared at three different sputtering time within 12 s using radio frequency (r.f.) magnetron sputtering and characterized by atomic force microscopy. Good nucleation uniformity has been obtained on the unabraded mica substrates due to the statistical distribution of nucleation sites. Especially, we have already observed an interesting unusual nucleation phenomenon, the island–rim structure on the abraded mica substrates after 4 s, where the island is fractal-like and the rim around the island was only occupied by few grains for nucleation. These ramified islands were located at the wide grooves introduced as predominate nucleation sites. The island–rim structure formed in initial nucleation process is closely associated with x, y velocity components of surface atoms and the distribution of active sites (related intimately to the surface free energy σ and strain energy ε) for nucleation. With the increasing of the sputtering time, voids and overlarge grains have been observed in the island–rim region after the sputtering time of 8 s and 12 s, respectively.