Photoluminescence (PL) of Fe3O4 nanoparticle was observed from the visible to near-infrared (NIR) range by laser irradiation at 407 nm. PL spectra of ∼10 nm diameter Fe3O4 nanoparticles organized in different spatial configuration, showed characteristic emissions with a major peak near 560 nm, and two weak peaks near 690 nm and 840 nm. Different band gap energies were determined for these Fe3O4 nanoparticle samples corresponding to, respectively, the electron band structures of the octahedral site (2.2 eV) and the tetrahedral site (0.9 eV). Photothermal effect of Fe3O4 nanoparticles was found to be associated with the photoluminescence emissions in the NIR range. Also discussed is the mechanism responsible for the photothermal effect of Fe3O4 nanoparticles in medical therapy.