AbstractWe report a multimodal biomicroscopic system which offers high-frequency ultrasound B-mode, acoustic radiation force impulse (ARFI), and multispectral imaging for qualitative tumor characterization ex vivo. Examinations of resected tissues from diseased regions such as tumors are crucial procedures during surgical operations to treat cancer. Particularly, if tiny tumors remain at surgical sites after tumor resection, such tumors can result in unwanted outcomes, such as cancer recurrence or metastasis to other organs. To avoid this, accurate characterizations of tumors resected during surgery are necessary. To this end, we devised a multimodal biomicroscopic system including high-frequency ultrasound B-mode, ARFI, and multispectral imaging modalities to examine resected tumors with high levels of accuracy. This system was evaluated with tissue-mimicking phantoms with different mechanical properties. In addition, colorectal tumors excised from cancer patients were examined. The proposed system offers highly resolved anatomical, mechanical, chemical information pertaining to tumors, thus allowing the detection of tumor regions from the surface to deep inside tissues. These results therefore suggest that the multimodal biomicroscopic system has the potential to undertake qualitative characterizations of excised tumors ex vivo.