CHI-KUANG SUNKao, Chien-TingChien-TingKaoWei, Ming-LiangMing-LiangWeiYI-HUA LIAO2019-12-042019-12-0420190277-786Xhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85072617502&doi=10.1117%2f12.2520417&partnerID=40&md5=81f40ba87a0dffc5daa4d260bf28c21ahttps://scholars.lib.ntu.edu.tw/handle/123456789/434780Intraoperative 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. ? 2019 SPIE. Downloading of the abstract is permitted for personal use only.enCr:forsterite; Eosin; Hematoxylin; Histopathology; Mohs surgery; Nonlinear microscopy; Third harmonic generation; Three-photon fluorescence[SDGs]SDG3Chromium compounds; Fluorescence; Histology; Multiphoton processes; Nonlinear optics; Olivine; Pathology; Photons; Surgery; Tissue; Cr: forsterite; Eosin; Hematoxylin; Histopathology; Mohs surgery; Nonlinear microscopy; Three-photon; Harmonic generationconference paper10.1117/12.25204172-s2.0-85072617502