Mechanical micromachining using micro-machine tools is very competitive in manufacturing geometrically complex 3D micro-parts in various engineering materials with advantages of small space, energy-efficient, low equipment production and operation costs. As key components of micro-machine tools, micro-spindles directly determine the performance of micro-machine tools and the application and development of mechanical micromachining technology. This study proposes a novel design concept of the separately spindle and tool and one-piece tool/rotor structure for micro-spindles, whose micro-tool can maintain good rotational accuracy at high rotational speeds. In the principle prototype micro-spindle, an air driven turbine was used as power spindle. The design and optimization of the micro-spindle were conducted. The fabrication and evaluation of the prototype micro-spindle were presented. The results of the principle prototype micro-spindle were analyzed and some efforts for the subsequent micro-spindle were proposed.