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

Yogesh G. Patel; Kishwer S. Nehal; Allan C. Halpern; Milind Rajadhyaksha

doi:10.1117/12.589394

25 April 2005 Confocal reflectance microscopy of basal cell cancers ex vivo: progress toward enhancing contrast and detectability of nuclei relative to dermis

Yogesh G. Patel, Kishwer S. Nehal, Allan C. Halpern, Milind Rajadhyaksha

Author Affiliations +

Proceedings Volume 5686, Photonic Therapeutics and Diagnostics; (2005) https://doi.org/10.1117/12.589394
Event: SPIE BiOS, 2005, San Jose, CA, United States

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.

Citation Download Citation

Yogesh G. Patel, Kishwer S. Nehal, Allan C. Halpern, and Milind Rajadhyaksha "Confocal reflectance microscopy of basal cell cancers ex vivo: progress toward enhancing contrast and detectability of nuclei relative to dermis", Proc. SPIE 5686, Photonic Therapeutics and Diagnostics, (25 April 2005); https://doi.org/10.1117/12.589394

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available