Abstract: Objective Expressing GCaMP6f in nucleus accumbens of mice using brain stereotaxic injection，examined neuron activity of restrained mice by calcium imaging in real time. Methods To get recombination virus, cultured 293T cell, then packed and purified aav-Syn1-GCaMP6f-P2A- Tomato virus. After being confirmed the recombination virus worked by infecting primary neurons, the virus was injected into nucleus accumbens by stereotaxic injection. And after the animals recovered, the mice were implanted optical fiber in brains, and restrained, then recorded neuronal activity by calcium imaging in the nucleus accumbens in time. Results After stimulated by glutamic acid, the neurons expressed GCaMP6f protein released green fluorescence because of the calcium influx. Brain slices showed that the green fluorescence produced by GCaMP6f and the red fluorescence produced by a carried marker protein in virus could co-localize in the same neuron. Conclusions The adeno-associated virus aav- Syn1-GCaMP6f-P2A-Tomato could be successfully used to real timely monitor activated neurons employing calcium imaging in vivo.

Key words: Key words: calcium imaging, adeno-associated virus, nucleus accumbens, GCaMP6f