Slide-free histopathological imaging of hematoxylin-eosin-stained whole mount tissues using Cr:forsterite laser-based nonlinear microscopy

Chi-Kuang Sun; Chien-Ting Kao; Ming-Liang Wei; Yi-Hua Liao

doi:10.1117/12.2520417

30 April 2019 Slide-free histopathological imaging of hematoxylin-eosin-stained whole mount tissues using Cr:forsterite laser-based nonlinear microscopy

Chi-Kuang Sun, Chien-Ting Kao, Ming-Liang Wei, Yi-Hua Liao

Author Affiliations +

Proceedings Volume 11026, Nonlinear Optics and Applications XI; 110260T (2019) https://doi.org/10.1117/12.2520417
Event: SPIE Optics + Optoelectronics, 2019, Prague, Czech Republic

Abstract

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.

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Citation Download Citation

Chi-Kuang Sun, Chien-Ting Kao, Ming-Liang Wei, and Yi-Hua Liao "Slide-free histopathological imaging of hematoxylin-eosin-stained whole mount tissues using Cr:forsterite laser-based nonlinear microscopy", Proc. SPIE 11026, Nonlinear Optics and Applications XI, 110260T (30 April 2019); https://doi.org/10.1117/12.2520417

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