The spindle is the main component in machine tools. The static and dynamic characteristics of the spindle directly affect the machining accuracy of workpieces. The characteristics of the spindle depend on the shaft size, bearing span, built-in motor location, and so on. Therefore, the appropriate selection of these parameters is important to improve the spindle characteristics. This paper presents the analysis of the static and dynamic characteristics and optimization design of a 40,000-rpm high-speed spindle. Statistical analysis for optimization and finite element analysis were performed. This study uses the response surface method to optimize the objective function and design factors. The targets are the natural frequency and displacement. The design factors are the shaft length, shaft diameter, bearing span, and motor location. The optimized design provides better results than the initial model, and these results are expected to improve the static and dynamic characteristics of the spindle.