Molecular Probes for Tracking Reactive Oxygen Species at the Organelle Interface

The spatially compartmentalized production and regulation of reactive oxygen species (ROS) play pivotal roles in cellular physiology and pathology. Over the past two decades, significant technological advances have propelled the development of fluorescent molecular probes and genetically encoded sensors to monitor redox dynamics within distinct organelles. This perspective review discusses recent progress and associated challenges in ROS tracking at the organelle interface, with particular emphasis on mitochondria, the endoplasmic reticulum (ER), lysosomes, and the emerging frontier of peroxisomal imaging. We examine chemical and protein‐based probe design strategies, target specificity issues, and the integration of multiplexed imaging techniques. Particular attention is given to the prospects for peroxisomal ROS tracking, a compartment historically underexplored despite its critical role in lipid metabolism and redox regulation and offer perspectives on how novel probe engineering may overcome current limitations. The ongoing evolution of these imaging modalities promises to deepen our understanding of subcellular redox signaling in both physiological and pathological states.