Significance: Recent research has identified key roles for reactive oxygen species (ROS)/reactive nitrogen species (RNS) in redox signaling, but much remains to be uncovered. Molecular imaging tools to study these processes must not only be selective to enable identification of the ROS/RNS involved but also reversible to distinguish signaling processes from oxidative stress. Fluorescent sensors offer the potential to image such processes with high spatial and temporal resolution.
Recent advances: A broad array of strategies has been developed that enable the selective sensing of ROS/RNS. More recently, attention has turned to the design of reversible small-molecule sensors of global redox state, with a further set of probes capable of reversible sensing of individual ROS/RNS.
Critical issues: In this study, we discuss the key challenges in achieving simultaneous detection of reversible oxidative bursts with unambiguous determination of a particular ROS/RNS.
Future directions: We have highlighted key design features of small-molecule probes that show promise in enabling the study of redox signaling, identifying essential parameters that must be assessed for any new probe. Antioxid. Redox Signal. 24, 713-730.