Nitric oxide (NO) is emerging as a very important and ubiquitous gaseous messenger in the body. The response characteristics of NO sensors made of non-conducting polymer modified carbon fiber electrodes are investigated to determine their selectivity, sensitivity, and stability for in vivo use. A composite polymer, comprising Nafion, m-phenylenediamine, and resorcinol, showed the best selectivity and stability to amperometric NO detection. The non-conducting, self-limiting polymer film protects the electrode from interference and fouling by other biochemicals. Although the relative sensitivity to NO of the modified sensor is lower than that of the unmodified carbon fiber electrodes (less than 6%), the composite polymer electrode showed high selectivity against ascorbic acid (> 2000:1), nitrite (> 600:1), and dopamine (> 200:1). The stability of the NO sensor was maintained for at least 1 week. The NO sensitivity after in vivo experiments (n = 8) is 88.1 +/- 5.6% of initial sensitivity data obtained before in vivo experiments. Preliminary in vivo experiments done with this electrode are shown to capture elevated NO levels in brain following an ischemic injury.