The application and optimization of a microsensor for in situ measurements of nitric oxide (NO) in biological systems are described. The sensor (diameter 0.5-0.8 mum), exhibiting a response time better than 10 ms and a detection limit of 10 nM, consists of several layers of p-type semiconducting polymeric porphyrin and cation exchanger (Nafion) deposited on a thermally-sharpened carbon fiber. The sensor has been applied to studies of NO release from a single endothelial cell in a pulmonary artery, as well as for the determination of NO in blood.