Confocal reflectance microscopy of basal cell cancers ex vivo: Progress toward enhancing contrast and detectability of nuclei relative to dermis Conference Paper

  

Authors:	Patel, Y. G.; Nehal, K. S.; Halpern, A. C.; Rajadhyaksha, M.
Editors:	Bartels, K. E.; Bass, L. S.; Riese, W. T. W.; Gregory, K. W.; Hirschberg, H.
Title:	Confocal reflectance microscopy of basal cell cancers ex vivo: Progress toward enhancing contrast and detectability of nuclei relative to dermis
Conference Title:	Photonic Therapeutics and Diagnostics
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.
Keywords:	cytology; basal cell carcinoma; confocal microscopy; skin cancer; histology; tumors; diagnosis; surgery; mohs surgery; acetowhitening; acetic acid; tissue; cells; imaging techniques; optical imaging; microscopic examination; backscattering
Journal Title	Proceedings of SPIE
Volume:	5686
Conference Dates:	2005 Jan 22
Conference Location:	San Jose, CA
ISBN:	0277-786X
Publisher:	SPIE
Location:	San Jose, CA
Date Published:	2005-01-01
Start Page:	1
End Page:	6
Language:	English
DOI:	10.1117/12.589394
PROVIDER:	scopus
DOI/URL:	http://www.scopus.com/inward/record.url?eid=2-s2.0-23244451008&partnerID=40&md5=f3f36e0d3d10560b7778956e51e163b5
Notes:	--- - Progress in Biomedical Optics and Imaging - Proceedings of SPIE - Progr. Biomed. Opt. Imaging Proc. SPIE - "Conference code: 65350" - "Cited By (since 1996): 1" - "Export Date: 24 October 2012" - "Art. No.: 01" - "Sponsors: SPIE" - 22 January 2005 through 25 January 2005 - "Source: Scopus"

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