•Multi-frequency ultrasound assisted freezing accelerated the freezing rate.•Increase in the number of ultrasonic frequencies enhanced the cavitation effects.•TUF treated samples retained better quality attributes.•Light microscopy observation was adopted to study the microstructures of frozen fish. This research evaluated the effects of multi-frequency ultrasound assisted freezing (UAF) on the freezing rate, structural characteristics, and quality properties of cultured large yellow croaker. The freezing effects with triple ultrasound-assisted freezing (TUF) at 20, 28 and 40 kHz under 175 W was more obvious than that of single ultrasound-assisted freezing (SUF) at 20 kHz and dual ultrasound-assisted freezing (DUF) at 20 and 28 kHz. The results showed that UAF significantly increased the freezing rate and better preserved the quality of frozen large yellow croaker samples. Specifically, the quality parameters of the TUF-treated samples were closer to those of the fresh samples, with greater texture characteristics, a larger water holding capacity (lower thawing loss and cooking loss), lower K values and lower thiobarbituric acid reactive substances values. Light microscopy observation images revealed that the ice crystals formed by TUF were fine and evenly distributed, resulting in less damage to the frozen large yellow croaker samples. Therefore, multi-frequency UAF could improve the quality properties of the large yellow croaker samples.