The total workflow lasts 2 days. This duration accepts some variation. Cells can be stored for a few days in PBS after fixation at 4 °C. We recommend using fresh cells whenever possible but cryopreserved cells in fetal calf serum/10% DMSO could also be used. Moreover, before image acquisition, the stained cells can be kept for a few days in the dark at 4 °C before the fluorescent signal decreases.