The current calibration methods of broadband ADCP cannot effectively realize the measurement of flow rate stratification. An acoustic response calibration method based on frequency shift processing by signal resampling is proposed in this paper. This method utilizes a group of response transducers to receive the signals transmitted by each transducer of the ADCP to be calibrated, then some simulated echo signals which contain Doppler shift information are generated by the process of signal segmentation and time-window compensation. In order to be consistent with the real water scattering situation, a calculation model of the echo signal amplitude is given based on the active sonar equation. A broadband ADCP calibration system for multi-cell is established, which could be applied in onland and underwater, and two ADCP devices are calibrated by designing some experiments. The measurement uncertainty of the calibration system is less than 0. 3% v+5 mm/ s (v denotes the flow velocity) (k = 2) in the flow velocity range of 0. 01 ~ 10 m/ s. The proposed method and calibration system are also verified by comparing to the flume towing method through experiment. Keywords:acoustic response calibration method; signal resampling; flow rate stratification; time window compensati