ADCPs have been widely used to estimate the dynamic characteristics and biomass of sound scattering layers (SSLs), and swimming speed of fish schools for analyzing SSLs spatial distribution and/or various behavior patterns. This result showed that the verification of the mean volume backscattering strength (MVBS or averaged SV, dB) acquired by the ADCP would be necessary for a quantitative analysis on the spatial distribution and the biomass estimation of the SSLs or fish school when ADCP is used for estimating their biomass. In addition, the calibrated sphere method was used to verify values of each MVBS obtained from 4 beams of ADCP (153.6 kHz) on the base of 3 frequencies (38, 120, 200 kHz) of Scientific echo sounder's split beam system. Then, the measured SV values were compared and analyzed in its Target Strength (TS, dB) values estimated by a theoretical acoustic scattering model.