Gold nanorods (GNRs) are a unique tool for imaging and tracking single particles in live cells, due to their excellent brightness, high stability, compatibility with cellular conditions and easy functionalization. We use a two-photon multifocal scanning microscope to acquire 3D movies of cells with high spatial and temporal resolution. Non-functionalized GNRs were delivered in the cytoplasm and nucleus of mammalian cells via microinjection in single cell. We followed more than 1000 GNRs trajectories going from few seconds to over 20 minutes. GNRs show similar behavior in the nucleus and in the cytoplasm. 90% are confined, yielding a diffusion coefficient of 0.05 µm2/s and a confinement smaller than 0,5 μm, and 10% appear to undergo directed motion with a velocity of 0.02 μm/s. Simulations of diffusing particles were used to test the accuracy of the results, and optimize the acquisition and analysis process. We aim to functionalize GNRs with proteins to study single protein-DNA interactions in the nucleus.