Aiming at the problem of small number of suspended particles and limited levitation space in the current three-dimensional acoustic operation technology, a design scheme of acoustic levitation device composed of three pairs of opposed transducer arrays is proposed. By changing the phase difference of the transducer array, the three-dimensional manipulation of particles is successfully realized. This device can be used to measure the speed of sound with a relative error generally about 2%~5%, and the measurement should carry out under a relatively strict environment.