We develop a stereo-multispectral endoscopic prototype in which a filter-wheel is used for surgical guidance to remove cholesteatoma tissue in the middle ear. Cholesteatoma is a destructive proliferating tissue. The only treatment for this disease is surgery. Removal is a very demanding task, even for experienced surgeons. It is very difficult to distinguish between bone and cholesteatoma. In addition, it can even reoccur if not all tissue particles of the cholesteatoma are removed, which leads to undesirable follow-up operations. Therefore, we propose an image-based method that combines multispectral tissue classification and 3D reconstruction to identify all parts of the removed tissue and determine their metric dimensions intraoperatively. The designed multispectral filter-wheel 3D-endoscope prototype can switch between narrow-band spectral and broad-band white illumination, which is technically evaluated in terms of optical system properties. Further, it is tested and evaluated on three patients. The wavelengths 400 nm and 420 nm are identified as most suitable for the differentiation task. The stereoscopic image acquisition allows accurate 3D surface reconstruction of the enhanced image information. The first results are promising, as the cholesteatoma can be easily highlighted, correctly identified, and visualized as a true-to-scale 3D model showing the patient-specific anatomy.