A surfactant-assisted method for preparing colloidal spheres with narrow size distribution from a polydispersed azo polymer has been developed in this work. The colloidal spheres were formed through gradual hydrophobic aggregation of the polymeric chains in THF-H2O dispersion media, which was induced by a steady increase in the water content. Results showed that the addition of a small amount of surfactant (SDBS) could significantly narrow the size distribution of the colloidal spheres. The size distribution of the colloidal spheres was determined by the concentrations of azo polymer and the amount of surfactant in the systems. When the concentrations of polymer and surfactant amount were in a proper range,colloidal spheres with narrow size distribution could be obtained. The colloidal spheres formed by this method could be elongated along the polarization direction of the laser beams upon Ar+ laser irradiation. The colloidal spheres are considered to be a new type of the colloid-based functional materials.