Cumulative evidence suggests that cellular senescence plays a variety of important physiological roles, including tumor suppression, embryonic development and ageing. Senescent cells are characterized by increased production of reactive oxygen species (ROS), mostly produced by dysfunctional mitochondria. Both intracellular and extracellular ROS have been shown to contribute to the induction of senescence. ROS have also been shown to act as signaling molecules during senescence, stabilizing the cell-cycle arrest. In this chapter, we present a detailed description of protocols that allow us to characterize intracellular and extracellular ROS in live senescent cells.