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Proceedings Paper

Confocal reflectance microscopy of basal cell cancers ex vivo: progress toward enhancing contrast and detectability of nuclei relative to dermis
Author(s): Yogesh G. Patel; Kishwer S. Nehal; Allan C. Halpern; Milind Rajadhyaksha
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Paper Abstract

Mohs surgery is a staged procedure for microscopically excising basal cell carcinomas (BCCs) while preserving the surrounding normal skin. Serial excisions are performed with each excision being guided by examination of the frozen histology. Mohs surgery is a meticulous and time-consuming (15-45 minutes per excision) procedure requiring several (2-20) excisions and frozen histology prepared for each excision. Real-time confocal reflectance microscopy may make Mohs surgery more efficient by enabling rapid detection of BCCs directly in fresh, unprocessed excisions, and thereby possibly avoiding frozen histology. As previously reported, we are developing an acetowhitening-and-cross polarized method to detect BCCs with a confocal reflectance microscope. Acetowhitening compacts the chromatin within the nucleus, increasing nuclear backscatter, and brightening the nuclei in the confocal images of the tissue. Our experiments to optimize acetowhitening, using acetic acid concentrations from 1% to 30% and treatment times from 30 seconds to 5 minutes, show that a minimum concentration of 2% with minimum washing time of 2 minutes is required for enhancing nuclear morphology. Increased depolarization is observed within the compacted chromatin relative to the surrounding collagen, and imaging in brightfield or crossed polarization brightens or darkens the cellular cytoplasm and birefringent dermis; thus, we may potentially vary nuclear/cytoplasm and nuclear/dermis contrast. Images are collected, oriented, and tiled to create mosaics and sub-mosaics to view large excisions at variable 2X - 10X magnifications. To create and display mosaics, adequate pixelation relative to resolution must be maintained and precise mechanical fixturing is necessary to control tilt, sag, flattening and stability of the excised tissue specimen.

Paper Details

Date Published: 25 April 2005
PDF: 6 pages
Proc. SPIE 5686, Photonic Therapeutics and Diagnostics, (25 April 2005); doi: 10.1117/12.589394
Show Author Affiliations
Yogesh G. Patel, Memorial Sloan-Kettering Cancer Ctr. (United States)
Kishwer S. Nehal, Memorial Sloan-Kettering Cancer Ctr. (United States)
Allan C. Halpern, Memorial Sloan-Kettering Cancer Ctr. (United States)
Milind Rajadhyaksha, Memorial Sloan-Kettering Cancer Ctr. (United States)
Northeastern Univ. (United States)

Published in SPIE Proceedings Vol. 5686:
Photonic Therapeutics and Diagnostics
Brian Jet-Fei Wong M.D.; Eugene A. Trowers M.D.; Kenton W. Gregory M.D.; Abraham Katzir; Nikiforos Kollias; Reza S. Malek M.D.; Henry Hirschberg M.D.; Kenneth Eugene Bartels D.V.M.; Steen J. Madsen; Lloyd P. Tate V.D.M.; Lawrence S. Bass M.D.; Werner T. W. de Riese; Karen M. McNally-Heintzelman, Editor(s)

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