Based on the auto-fluorescence property of collagen in articular cartilage, the research on fluorescence microscopy measurement method of articular cartilage was carried out. First, the difference between the healthy and pathological samples was revealed by statistical comparison of fluorescence images. Second, based on the polarization property of fluorescence, the fluorescence anisotropy of the cartilage was presented by the linear dichroism method, which lays foundation for research on structure of articular cartilage using fluorescence anisotropy method. Compared to traditional microscopic measurement techniques that supply the morphology and the super-structure of cartilage tissues, fluorescence microscopy measurement methods are able to obtain the information about the structure of cartilage tissues. The results presented in this paper open the door for developing new diagnostic method for articular cartilage diseases.