SPT allowed observing moving particles. These trajectories are used to investigate cytoplasm or membrane organization, but also the cell nucleus dynamics, remodeler dynamics or mRNA production. Due to the constant improvement of the instrumentation, the spatial resolution is continuously decreasing, reaching now values of approximately 20 nm, while the acquisition time step is usually in the range of 10 to 50 ms to capture short events occurring in live tissues. A variant of super-resolution microscopy called sptPALM is used to detect the local and dynamically changing organization of molecules in cells, or events of DNA binding by transcription factors in mammalian nucleus. Super-resolution image acquisition and particle tracking are crucial to guarantee a high quality data