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A new spot centroid detection algorithm for a Shack-Hartmann wavefront sensor (SHWFS) is experimentally investigated. The algorithm is a kind of dynamic tracking algorithm that tracks and calculates the corresponding spot centroid of the current spot map based on the spot centroid of the previous spot map, according to the strong correlation of the wavefront slope and the centroid of the corresponding spot between temporally adjacent SHWFS measurements. That is, for adjacent measurements, the spot centroid movement will usually fall within some range. Using the algorithm, the dynamic range of an SHWFS can be expanded by a factor of three in the measurement of tilt aberration compared with the conventional algorithm, more than 1.3 times in the measurement of defocus aberration, and more than 2 times in the measurement of the mixture of spherical aberration plus coma aberration. The algorithm is applied in our SHWFS to measure the distorted wavefront of the human eye. The experimental results of the adaptive optics (AO) system for retina imaging are presented to prove its feasibility for highly aberrated eyes.