Conjugated copolymers containing fluorene and diphenylamine moieties with octyl and triethylene glycol side chains have been synthesized via Suzuki-Miyaura cross-coupling reaction and their structures have been characterized. Conjugated polymer nanoparticles and self-organization of the polymers have been prepared by rapid precipitation and solvent diffusion methods, respectively. The conjugated polymers exhibited well-defined spherical amorphous structures and crystalline microspheres with a diameter ranging from several nanometer to micrometer. Self-assembled conjugated polymer aggregates were obtained in a selective good solvent/poor solvent mixture. Alternating conjugated polymers are generally difficult to assemble into well-defined spheres due to their rigid and planar backbones, however, alternating polymers containing dioctyl and triethylene glycol side chains tends to form microspheres or sheet-like structures depending on the different solvent mixtures. The emission maximum of the dispersed polymers in water was significantly red-shifted with a dramatic reduction in the photoluminescence quantum yield. In addition, the emission maximum of the self-assembled copolymers was barely red shifted with a small reduction in the photoluminescence quantum yield.