Label-free and non-destructive histology of unprocessed biological tissues with ultraviolet single-plane illumination microscopy

Abstract

Histopathology has remained the gold standard for surgical margin assessment for decades. However, routine pathological examination based on formalin-fixed and paraffin-embedded (FFPE) tissues is laborious and time-consuming, failing to guide surgeons intraoperatively. Here, we propose a rapid, label-free, and non-destructive histological imaging method, termed microscopy with ultraviolet single-plane illumination (MUSI). With intrinsic fluorescence from deep ultraviolet excitation, MUSI enables both ex vivo and in vivo imaging of fresh and unprocessed tissues at the subcellular level with an imaging speed of 0.5 mm2/s, generating high-quality optically sectioned surface images from irregular surgical tissues with a long depth-of-field. We demonstrate that MUSI could differentiate between different subtypes of human lung adenocarcinomas (e.g., lepidic, acinar, papillary, and micropapillary), revealing diagnostically important features that are comparable to the gold standard FFPE histology. As an assistive imaging platform, MUSI can provide immediate feedback to surgeons and pathologists for intraoperative decision-making, holding great promise to revolutionize the current clinical practice in surgical pathology.