Electrochemical nanosensor for real-time direct imaging of nitric oxide in living brain

As gaseous nitric oxide (NO), a critical and multifaceted biomarker, diffuses easily once released, identifying the precise sources of NO release is a challenge. This study developed a new technique for real-time in vivo direct NO imaging by coupling an amperometric NO nanosensor with scanning electrochemical microscopy. This technique provides three-dimensional information of the NO releasing sites in an intact living mouse brain with high sensitivity and spatial resolution. Immunohistochemical analysis was carried out to confirm the anatomical reliability of the acquired electrochemical NO image. The real-time NO imaging results were well matched with the corresponding immunohistochemical analysis of neuronal NO synthase immunoreactive (nNOS-IR) cells, i.e., NO releasing sites in a living brain. The imaged NO local concentrations were confirmed to be closely related to the location in depth, the size of the nNOS-IR cell, and the intensity of nNOS immunoreactivity. This paper demonstrates the first direct electrochemical NO imaging of a living brain.