A self-built micro-particle image velocimetry (micro-PIV) with a diode laser is established to measure the micro-fluidic phenomenon in a 100 μm rectangular capillary. By scanning method, a 3-D flow image with a flowrate of 0.3 μL/min is presented. With this calibration method, the measurement ability for 3-D micro-fluidic dynamics could be achieved. This technique also reveals its benefit and potential in metrology. Hence, it provides a helpful tool for Bio-MEMS research. The experiment is proceeded under laminar flow, Re= 0.011. The measurement range is ranging from 0.05μm/s to 4.3mm/s. The vector grid resolution is optimized to 2.5 μm.