A dual-channel macroscopic imaging system has been developed based on optical methods， which can simultaneously capture the visible light and near-infrared second window （NIR-II） fluorescence. It could provide high-quality bright-field real-time images with the NIR-II fluorescence information， addressing the significant disparity issue between fluorescence and bright-field images in conventional NIR-II macroscopic imaging systems. In the experiment， the anti-scattering capability and imaging performance of NIR-II fluorescence signals of indocyanine green （ICG） were tested using different thicknesses of adipose tissues in the band of 1 100-1 700 nm and 1 300-1 700 nm respectively. Subsequently， the dual-channel macroscopic imaging system was used to obtain lymph node images of mouse and rat models， simulating the lymph node resection surgery and mimicking the process of abdominal lymph node clearance. Finally， different thicknesses of biological adipose tissues were added to the rat model to simulate the presence of adipose tissues covering the lymph nodes during actual surgery， and the penetration capability of the dual-channel system was observed. The visible and near-infrared second window dual-channel fluorescence imaging system provided the intuitive visual information to the operator， reducing surgery time and improving the patient prognosis， and held great potential for application in clinical surgical navigation.