The invention discloses a multistage thermoelectric cooler parameter vacuum testing device which comprises a vacuum cavity, a vacuum pump, a water-cooling head, a temperature control cooling water circulation device, a heating resistor module, a cold-end temperature measurement resistor, a hot-end temperature measurement resistor, a direct current power source, and a data processing and displaying module. A vacuum pumping valve and an air vent valve are arranged on the vacuum cavity. The vacuum cavity is sealed with the water-cooling head through a vacuum rubber ring and connecting screws. A hot end of a thermoelectric cooler achieves heat dissipation and temperature control through the water-cooling head and the temperature control cooling water circulation device. The data processing and displaying module is used for carrying out real-time control, data processing and result displaying on appliances in the device. Through adoption of the multistage thermoelectric cooler parameter vacuum testing device, the current-voltage response relation and the current-refrigerating capacity relation of the multistage thermoelectric cooler, the refrigerating efficiency of the multistage thermoelectric cooler under different heat dissipation conditions and other parameters can be obtained. The multistage thermoelectric cooler parameter vacuum testing device is simple in structure, convenient to operate, high in measuring accuracy, and capable of effectively preventing a cold end of the thermoelectric cooler from dewing in the testing process of large temperature difference.