Fluorescence confocal laser scanning endomicroscopy is a novel tool combining confocal microscopy and endoscopy for in-vivo subcellular structure imaging with comparable resolution as the traditional microscope. In this paper, we propose a three-channel fluorescence confocal microscopy system based on fiber bundle and two excitation laser lines of 488nm and 650nm. Three fluorescent photomultiplier detecting channels of red, green and blue can record multi-color fluorescence signals from single sample site simultaneously. And its ability for in-vivo multi-channel fluorescence detection at subcellular level is verified. Moreover, the system has achieved an effective field of view of 154μm in diameter with high resolution. With its multi-laser scanning, multi-channel detection, flexible probing, and in-vivo imaging abilities it will become a powerful tool in bio-chemical research and diagnostics, such as the investigation of the transport mechanism of nano-drugs in small animals.
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