The invention discloses a detachable three-electrode battery testing device and an assembling method thereof. The device comprises a shell, a lead screw, an extrusion assembly and a reference electrode assembly, wherein the shell comprises a base, a first sealing ring and an upper cover; a positioning groove is formed in the cavity bottom of the base; the upper cover is detachably connected to the top end of the base; the first sealing ring is arranged between the upper cover and the base; the lead screw penetrates through the upper cover and the first sealing ring to be in threaded connection with the base. The device can test and research the impedance and the change condition of the electrode plate, can research and analyze the dynamic mechanism in the chemical process of the electrode, can judge and analyze the battery capacity recession reason, is high in sealing performance, avoids air leakage and liquid leakage, can realize thousands of cycles of a power or energy storage battery, improves labor efficiency, the device is simple in structure, reasonable in arrangement, low in cost and high in practicability, and the assembling method is safe, few in step and easy to operate.