The genetic algorithm (GA) based stereo particle-pairing algorithm has been developed and applied to the spatial particle-pairing problem of the stereoscopic three-dimensional (3-D) PTV system. In this 3 D PTV system, particles viewed by two (or more than two) stereoscopic cameras with a parallax have to be correctly paired at every synchronized time step. This is important because the 3-D coordinates of individual particles cannot be computed without the knowledge of the correct stereo correspondence of the particles. In the present study, the GA algorithm is applied to the epipolar line proximity analysis for establishing correspondence of particles pairs between two co-instantaneous stereoscopic particles images, in order to compute the 3-D coordinates of every individual particle. The results are tested with various standard images and it’s found that the new strategy using GA works better than conventional particle pairing methods of 3-D particle tracking velocimetry for steoroscopic PTV. Journal of the Institute of Engineering, 2016, 12(1): 10-26