A novel micro-photoelectrode with a selective molecular sieve was created for in vivo monitoring of O2 levels in the mouse brain. An ITO optical fiber modified by graphitized carbon nitride (g-C3N4) in situ was employed as the light activated substrate to provide rich photo-induced electrons for the catalytic reduction of O2. Meanwhile, the porous hybrid layer composed of zeolitic imidazolate framework-8 and polysulfone was constructed over the g-C3N4 surface as the molecular sieve to synergically enhance the selectivity of O2 detections. By advantage of this useful tool, the real time variation of the O2 level was successfully determined in the mouse brain upon ischemia.