Label-free, High-Resolution Optical Metabolic Imaging of Human Cervical Precancers Reveals Potential for Intraepithelial Neoplasia Diagnosis

While metabolic changes are considered a cancer hallmark, their assessment has not been incorporated in the detection of early or precancers, when treatment is most effective. Here, we demonstrate that metabolic changes are detected in freshly excised human cervical precancerous tissues using label-free, non-destructive imaging of the entire epithelium. The images rely on two-photon excited fluorescence from two metabolic co-enzymes, NAD(P)H and FAD, and have micron-level resolution, enabling sensitive assessments of the redox ratio and mitochondrial fragmentation, which yield metrics of metabolic function and heterogeneity. Simultaneous characterization of morphological features, such as the depth-dependent variation of the nuclear:cytoplasmic ratio, is demonstrated. Multi-parametric analysis combining several metabolic metrics with morphological ones enhances significantly the diagnostic accuracy of identifying high-grade squamous intraepithelial lesions. Our results motivate the translation of such functional metabolic imaging to in vivo studies, which may enable improved identification of cervical lesions, and other precancers, at the bedside. [Display omitted] Multiphoton microscopy enables label-free imaging of cervical epithelial biopsiesCell layer-dependent metabolic changes are detected in cervical precancersMorphofunctional multi-parametric evaluation enhances diagnostic accuracyIn vivo translation may enable noninvasive diagnostics at the bedside Reprogramming of cellular metabolism is an emerging cancer hallmark. Pouli et al. show that metabolic dysfunction can be detected in live human cervical biopsies using high-resolution autofluorescence images of the metabolic coenzymes NAD(P)H and FAD. The authors use morphological and functional metrics to successfully distinguish healthy tissues from precancerous lesions.