Endoscopic imaging systems are widely used in non-invasive diagnosis of internal tissues in biomedical applications. The imaging clarity and resolution of current fiber-optic endoscopes is far less than that of electronic endoscopes. However, the electronic endoscope has the loss of information related to the principle of the three primary colors of image transmission, which makes the color reproduction ability poor. In order to solve this problem, this paper proposes a hyperspectral imaging system that combines acousto-optic tunable filter (AOTF) and fiber endoscope. The system can obtain images at any wavelengths and obtain spectral information of samples. The system eliminates the loss of information related to the three-color principle. In this paper, firstly, a hyperspectral imaging system based on fiber endoscope is designed, then we test the performance of the system by imaging the resolution target, and finally evaluates the imaging and spectral resolution of the system. The experimental results show that the hyperspectral imaging system based on fiber endoscope has greater advantages than a single fiber endoscope system. The imaging resolution and image contrast of the system have been further improved, and the spectral bandwidth is about several nanometers. Any filtering center wavelength in the visible light range can be quickly selected through random access or continuous tuning, which provides more auxiliary information and convenience for medical diagnosis. The additional spectral information increases the reliability of medical diagnosis based on the data provided by the fiber endoscope. © 2021 COPYRIGHT SPIE.