Intraoperative margin assessment during Mohs micrographic surgery (MMS) is clinically critical. Currently, frozen pathology is the gold standard of assessing Mohs excisions for signs of remaining cancerous lesions; however, it is still considered time-consuming and sometimes misses the real margins. In this study, we demonstrate nonlinear microscopy imaging of hematoxylin-eosin (H&E) stained whole-mount skin tissues with a sub-femtoliter resolution through third-harmonic- generation (THG) and three-photon-excited-fluorescence (3PF) by using 1260 nm Cr:forsterite-laser-based nonlinear microscopy, which also enables virtual biopsy for preoperative margin assessment. Without the physical sectioning procedure and with a simplified staining process, real margins were preserved and time before microscopic examination was potentially saved three times shorter than frozen pathology. By exactly stained by H&E, the compatibility allows the adjunction to frozen pathology if further examination is needed. Virtual-sectioning imaging of H&E stained skin tissues is displayed real-time and can be post-processed analogously to conventional H&E histology, facilitating pathologists for diagnosis.

Date Published: 30 April 2019
PDF: 9 pages
Proc. SPIE 11026, Nonlinear Optics and Applications XI, 110260T (30 April 2019); doi: 10.1117/12.2520417

Published in SPIE Proceedings Vol. 11026:
Nonlinear Optics and Applications XI
Mario Bertolotti; Alexei M. Zheltikov, Editor(s)