In this paper, we report a novel organic fluorescent nanoparticle based on exciplexes for cell imaging. Through a reprecipitation method, we used a combination of 1,1-bis((di-4-tolylamino)phenyl)cyclohexane (TAPC) and 2,7-bis(diphenylphosphoryl)-9,9′-spirobi[fluorene] (SPPO13) to form nanoparticles. In the aggregated structures, TAPC and SPPO13 were forced into proximity that led to the corresponding exciplex formation. A red-shifted fluorescence emission with considerably longer fluorescence lifetimes ascribed to exciplex emission can be achieved. Along with the good stability and low cytotoxicity of organic nanoparticles, the prepared TAPC/SPPO13 exciplex nanoparticles were successfully applied in live cell imaging. These properties make TAPC/SPPO13 exciplex nanoparticles good candidates for cellular labeling and imaging materials.