Fluorescence imaging, in combination with
tumor-avid near-infrared (NIR) fluorescent
molecular probes, provides high specificity and sensitivity for
cancer detection in preclinical animal models, and more recently, assistance during oncologic surgery. However, conventional camera-based fluorescence imaging techniques are heavily surface-weighted such that surface reflection from skin or other nontumor tissue and nonspecific fluorescence signals dominate, obscuring true
cancer-specific signals and blurring
tumor boundaries. To address this challenge, we applied structured illumination fluorescence molecular imaging (SIFMI) in live animals for automated subtraction of nonspecific surface signals to better delineate accumulation of an NIR
fluorescent probe targeting α4β1
integrin in mice bearing subcutaneous plasma cell xenografts. SIFMI demonstrated a fivefold improvement in
tumor-to-background contrast when compared with other full-field fluorescence imaging methods and required significantly reduced scanning time compared with diffuse optical spectroscopy imaging. Furthermore, the spatial gradient mapping enhanced highlighting of
tumor boundaries. Through the relatively simple hardware and software modifications described, SIFMI can be integrated with clinical fluorescence imaging systems, enhancing intraoperative
tumor boundary delineation from the uninvolved tissue.