The primary goal of cancer surgery is to completely eliminate tumors. A real-time diagnostic method uses a fluorescence contrast agent and a surgical microscope to assess the status of tumor resection and the patient’s blood circulation. The biggest problem in imaging diagnostics using a microscope is the specular reflection phenomenon. While observing a lesion, the observation field may be obstructed due to specular reflection, making it difficult to obtain accurate results during the diagnostic process. Herein we propose a method to reduce specular reflection during tumor diagnosis by introducing a linearly polarized filter for a surgical microscope system. The method of angular direction adjustment of the filter ensures that only the horizontally polarized light passes through it, thereby obstructing the specular reflection. As a result of removing specular reflection, clear images were obtained at 90° and 270°. This experiment was conducted using phantoms and animals. Our results prove that the proposed method can be applied to imaging cameras used in internal medicine, surgery, and radiology for diagnosis.