Abstract: An electro-wetting-based microfluidic droplet positioning system, includes an electro-wetter, a microprocessor, a main control module, a droplet drive module, a droplet positioning module and a power supply. Further, an electrowetting-based microfluidic droplet positioning method, includes the steps of: considering, by a system, a droplet to be measured in an electro-wetter and a hydrophobic insulation layer below the droplet as a capacitor connected in series; issuing, by a main control chip, a command to a droplet drive module, and driving, by the droplet drive module, the droplet to be measured to move; collecting, by a droplet positioning module, a current capacitance value of the droplet, and determining a relative position of the droplet; and verifying, by the system, whether the droplet is at a target position.