At the nanoscale, the ZnGa2O4 spinel doped with chromium (III) is an interesting material for in vivo optical imaging due to its bright red persistent luminescence after UV and visible excitation. Moreover its persistent luminescent properties can be improved with the incorporation of bismuth (III) as a co-dopant without any structure changes. The nanoparticles are synthesized by soft chemistry using microwave heating in aqueous media. These very small sized nanophosphors (around 10 nm) present interesting long lasting persistent luminescence after annealing at 1000°C and they can be excited both under UV and under visible LED excitation. In this work we try to understand the mechanisms of the persistent luminescent properties of such nanomaterials. Thermoluminescence is performed to investigate trapping and detrapping processes as well as trap distribution. The chromium local environment is studied by Electron Paramagnetic Resonance. 71Ga Nuclear Magnetic Resonance is used to get information on the gallium ions repartition (tetrahedral or octahedral site) in the structure. Comparison of optical properties versus local structure increases the understanding of the persistent luminescence mechanism and gives insights to the new modalities for their use as nanoprobes for in vivo imaging.