A carbon fiber microelectrode modified with a composite film of carbon nanotubes and Nafion was developed for in vivo ascorbate (AA) measurements in brain tissue. The modified-microelectrodes exhibit an electrocatalytic activity for AA oxidation by shifting the peak potential negatively to À0.040 V, showing a sensitivity of 37 pA/mM, a detection limit of 2.5 mM, a response time of 0.3 s and dont respond to several electroactive compounds found in the brain extracellular space. In the rat hippocampus, the basal concentration of AA was 290 mM, and glutamateevoked changes in AA were biphasic comprising fast and slow components.