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Input coupling is an important factor needing to be considered in the process of designing a precision micro-positioning mechanism. Couplings between the input bodies will cause serious difficulties in the motion control. Firstly, an input coupling analysis method is presented for a 3-DOF micro-positioning stage. The method is validated by the ANSYS simulation and experiment results. Secondly, the influences of the structural parameters of the mechanism on the input coupling are discussed. Finally, three different contact cases versus the input coupling are analyzed, and an optimal design method is developed. Numerical results show that the input couplings of the optimized stage are reduced remarkably. The research results of this paper are helpful to design a low coupling micro-positioning system.