Comparison of nonlinear microscopy and frozen section histology for imaging of Mohs surgical margins
- 30 July 2019
- journal article
- research article
- Published by Optica Publishing Group in Biomedical Optics Express
- Vol. 10 (8), 4249-4260
- https://doi.org/10.1364/boe.10.004249
Abstract
Mohs surgery uses err force frozen section analysis (FSA) with complete margin examination for the excision of select basal cell carcinomas (BCC), obtaining excellent cosmetic outcomes and extremely low recurrence rates. However, Mohs with FSA is time-consuming because of the need to iteratively perform cryosectioning on sequential excisions. Fluorescent microscopies can image tissue specimens without requiring physical sectioning, potentially reducing the time to perform Mohs surgery. We demonstrate a protocol for nonlinear microscopy (NLM) imaging of surgical specimens that combines dual agent staining, virtual H&E rendering, and video rate imaging. We also introduce a novel protocol that enables micron-level co-registration of NLM images with FSA histology, and demonstrate that NLM can reproduce similar features similar to FSA in BCC specimens with both negative and positive surgical margins. We show that the fluorescent labels can be extracted with conventional vacuum infiltration processing, enabling subsequent immunohistochemistry on fluorescently labeled tissue. This protocol can also be applied to evaluate the performance of NLM compared with FSA in a wide range of pathologies for intraoperative consultation. (C) 2019 Optical Society of America under the terms of the OSA Open Access Publishing AgreementKeywords
Funding Information
- National Institutes of Health (NIH) (R01-CA075289, R01-CA178636)
- Air Force Office of Scientific Research (AFOSR) (FA9550-15-1-0473)
This publication has 19 references indexed in Scilit:
- Rapid histopathological imaging of skin and breast cancer surgical specimens using immersion microscopy with ultraviolet surface excitationScientific Reports, 2018
- In Vivo Multiphoton Microscopy of Basal Cell CarcinomaJAMA Dermatology, 2015
- Incidence Estimate of Nonmelanoma Skin Cancer (Keratinocyte Carcinomas) in the US Population, 2012JAMA Dermatology, 2015
- Evaluating ex vivo fluorescence confocal microscopy images of basal cell carcinomas in M ohs excised tissueBritish Journal of Dermatology, 2014
- Confocal microscopy with strip mosaicing for rapid imaging over large areas of excised tissueJournal of Biomedical Optics, 2013
- Mohs Micrographic Surgery and Surgical Excision for Nonmelanoma Skin Cancer Treatment in the Medicare PopulationArchives of Dermatology, 2012
- Confocal mosaicing microscopy of human skin ex vivo: spectral analysis for digital staining to simulate histology-like appearanceJournal of Biomedical Optics, 2011
- Detection of basal cell carcinomas in Mohs excisions with fluorescence confocal mosaicing microscopyBritish Journal of Dermatology, 2009
- Confocal mosaicing microscopy in Mohs skin excisions: feasibility of rapid surgical pathologyJournal of Biomedical Optics, 2008
- Basal cell carcinoma treated with Mohs surgery in Australia II. Outcome at 5-year follow-upJournal of the American Academy of Dermatology, 2005