We conducted an initial feasibility study using real-time magneto-motive optical Doppler tomography (MM-ODT) with enhanced contrast to investigate the detection of superparamagnetic iron oxide (SPIO) magnetic nanoparticles implanted into in vivo melanoma tissue. The MM-ODT signals were detected owing to the phase shift of the implanted magnetic nanoparticles, which occurred due to the action of an applied magnetic field. An amplifier circuit-based solenoid was utilized for generating high-intensity oscillating magnetic fields. The MM-ODT system was confirmed as an effective in vivo imaging method for detecting melanoma tissue, with the performance comparable to those of conventional optical coherence tomography and optical Doppler tomography methods. Moreover, the optimal values of the SPIO nanoparticles concentration and solenoid voltage for obtaining the uppermost Doppler velocity were derived as well. To improve the signal processing speed for real-time imaging, we adopted multithread programming techniques and optimized the signal path. The results suggest that this imaging modality can be used as a powerful tool to identify the intracellular and extracellular SPIO nanoparticles in melanoma tissues in vivo.