Fluorescence imaging technology can dynamically monitor gene and cell changing in live animals in real-time, with advantages such as high sensitivity, high resolution, and non-invasion. In recent years, it has been widely used in tumor research, gene expression research, drug development research, etc. The imaging wavelength of traditional fluorescence imaging technology falls in the visible and near-infrared-I region. Due to the absorption and scattering effects of light propagation in biological tissues, and the inherent fluorescence of biological tissues, traditional fluorescence imaging techniques still have significant limitations in penetration depth and image signal-to-noise ratio. In this work, a highly integrated near-infrared-II (NIR-II, 900—1880 nm) small animal living fluorescence imaging system is developed by taking the advantages of NIR-II fluorescence imaging technology, such as low absorption, low scattering, and deep penetration depth in biological tissues. And a method of enhancing and correcting fluorescence image is proposed to optimize fluorescence images. In this work, the biological tissue simulation experiments and live animal experiments are conducted to test the performance and imaging effect of the system. The experimental results show that the system has the advantages of deep penetration depth, high signal-to-noise ratio, and high sensitivity. Combined with commercial indocyanine green reagents and aggregation-induced emission dyes, this system can monitor the distribution of blood vessels in real time and continuously monitor deep tissues and organs in mice, and conduct the dynamically monitoring research in living mice in a conscious state. This helps to promote tumor research and drug development research in the field of biomedical imaging to enter a new stage.