Fluorescence has been proved to be a versatile and useful tool for animal in vivo imaging. It has been used widely in biomedical research and drug discoveries. With the recent development of the technology and instruments, the 2-D imaging system has been developed to a 3-D imaging in vivo. 3-D imaging will provide researchers detailed spatial information of the targets and its relationship with animal organs. This paper introduces a new design of an in vivo 3-D multispectral fluorescent imaging system, including the design of hardware platform and the imaging algorithm based on binocular stereo vision theory. The working principle of the system, structure of the hardware and the system diagram are introduced. The algorithm of binocular stereo vision is utilized to complete camera calibration, stereo rectification, and 3D coordinates computing based on OpenCV.
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