Development of a Handheld Submersible Chemiluminescent Sensor: Quantification of Superoxide at Coral Surfaces

Reactive oxygen species (ROS) are produced via various photochemical, abiotic, and biological pathways. The low concentration and short lifetime of the ROS superoxide (O₂^•-) make it challenging to measure in natural systems. Here, we designed, developed, and validated a DIver-operated Submersible Chemiluminescent sensOr (DISCO), the first handheld submersible chemiluminescent sensor. The fluidic system inside DISCO is controlled by two high-precision pumps that introduce sample water and analytical reagents into a mixing cell. The resultant chemiluminescent signal is quantified by a photomultiplier tube, recorded by a miniature onboard computer and monitored in real time via a handheld underwater LED interface. Components are contained within a pressure-bearing housing (max depth 30 m), and an external battery pack supplies power. Laboratory calibrations with filtered seawater verified instrument stability and precision. Field deployment in Cuban coral reefs quantified background seawater-normalized extracellular superoxide concentrations near coral surfaces (0-173 nM) that varied distinctly with coral species. Observations were consistent with previous similar measurements from aquaria and shallow reefs using a standard benchtop system. In situ quantification of superoxide associated with corals was enabled by DISCO, demonstrating the potential application to other shallow water ecosystems and chemical species.