Laser Speckle Contrast Imaging (LSCI), which used coherent light, has fully been used for observing blood flow due to its non-invasive, non-contact acquisition method. Generally, LSCI system uses just a single wavelength for measurement. In this research, first, considering the biological characteristics of different reflection rates and absorption, we use two lasers at 633nm and 855 nm and two CCD cameras to build a microscopic LSCI system. Second, by using Spatial, Temporal Speckle Contrast Analysis methods and analysis with Beer–Lambert law, the microcirculation can be in vivo visualized and oxygenation can be observed. Such developed system can be further used for in vivo animal studies.