The invention discloses a back-portable visual in-vivo calcium imaging device, and belongs to the technical field of in-vivo calcium imaging. The device includes a device body, a spine holder and fixing screws; the device body includes a track pulley system, four universal wheels and a spine holder base, the track pulley system is composed of two mutually perpendicular metal rods, wherein the twomutually perpendicular metal rods include a horizontal track and a longitudinal track, and the bottom end of the longitudinal track is provided with a microscope fixing platform; and the spine holderis connected to the spine holder base of the device main body through the four fixing screws, and is located at the inner lower part of the device main body. According to the mouse back-portable in-vivo calcium imaging device provided by the invention, a mouse spine can be fixed and is not easy to move, the mouse spine can be combined with the device main body carrying an ultramicroscope, and thedevice has the advantages of clever design, a simple and reasonable structure, stable clamping, good flexibility, convenient use, and strong compatibility, and can make calcium signal recording methods more abundant.