Inside chloroplasts, several abiotic stresses (including drought, high light, salinity, or extreme temperatures) induce a reduction in CO2 assimilation rates with a consequent increase in reactive oxygen species (ROS) production, ultimately leading to leaf senescence and yield loss. Photo-oxidation processes should therefore be mitigated to prevent leaf senescence, and plants have evolved several mechanisms to either prevent the formation of ROS or eliminate them. Technology evolution during the past decade has brought faster and more precise methodologies to quantify ROS production effects and damage, and the capacities of plants to withstand oxidative stress. Nevertheless, it is very difficult to disentangle photo-oxidative processes that bring leaf defence and acclimation, from those leading to leaf senescence (and consequently death). It is important to avoid the mistake of discussing results on leaf extracts as being equivalent to chloroplast extracts without taking into account that other organelles, such as peroxisomes, mitochondria, or the apoplast also significantly contribute to the overall ROS production within the cell. Another important aspect is that studies on abiotic stress-induced leaf senescence in crops do not always include a time-course evolution of studied processes, which limits our knowledge about what photo-oxidative stress processes are required to irreversibly induce the senescence programme. This review will summarize the current technologies used to evaluate the extent of photo-oxidative stress in plants, and discuss their advantages and limitations in characterizing abiotic stress-induced leaf senescence in crops.