An electrochemical microsensor based on carbon ®ber microelectrode (CFME) chemically modi®ed with the per¯uorinated cationexchange polymer, Na®on and methylviologen (MV) and its application for in vivo voltammetric measurements of oxygen (O 2 ) are described. The microsensor shows a high catalytic activity for the reduction of O 2 with a good reproducibility, high sensitivity and selectivity and signi®cant ability against electrode fouling. The current is linear with the concentration of O 2 in a range from 9.0610 À6 to 2.0610 À4 molyL with a calculated detection limit, at a signal-to-noise ratio of 3 to 1, of 5.0610 À6 molyL and correlation coef®cient of 0.9985. The relative standard deviation for 2.0610 À4 molyL O 2 is 1.7 % (n 12). Some compounds common to biological¯uids such as glucose, ascorbic acid, uric acid, catecholamine, glutamate, glutathione, Mg 2 , Ca 2 , Na , K and Cl À are tested in vitro and show no interferences with the voltammetric responses to O 2 . In vivo performance of the O 2 microsensors is demonstrated by measurements of local changes in O 2 in the brain of anesthetized rat before and during transient ischemia.