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Journal of Biomedical Optics

Tri-modal confocal mosaics detect residual invasive squamous cell carcinoma in Mohs surgical excisions
Author(s): Daniel S. Gareau; Anna Bar; Nicholas Snavely; Kenneth K. Lee; Neil Swanson; Eric Simpson; Steve Jacques; Nathaniel W. Chen
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Paper Abstract

For rapid, intra-operative pathological margin assessment to guide staged cancer excisions, multimodal confocal mosaic scan image wide surgical margins (approximately 1 cm) with sub-cellular resolution and mimic the appearance of conventional hematoxylin and eosin histopathology (H&E). The goal of this work is to combine three confocal imaging modes: acridine orange fluorescence (AO) for labeling nuclei, eosin fluorescence (Eo) for labeling cytoplasm, and endogenous reflectance (R) for marking collagen and keratin. Absorption contrast is achieved by alternating the excitation wavelength: 488 nm (AO fluorescence) and 532 nm (Eo fluorescence). Superposition and false-coloring of these modes mimics H&E, enabling detection of cutaneous squamous cell carcinomas (SCC). The sum of mosaic Eo+R is false-colored pink to mimic the appearance of eosin, while the AO mosaic is false-colored purple to mimic the appearance of hematoxylin in H&E. In this study, mosaics of 10 Mohs surgical excisions containing invasive SCC, and five containing only normal tissue were subdivided for digital presentation equivalent to 4× histology. Of the total 50 SCC and 25 normal sub-mosaics presented, two reviewers made two and three type-2 errors (false positives), respectively. Limitations to precisely mimic H&E included occasional elastin staining by AO. These results suggest that confocal mosaics may effectively guide staged SCC excisions in skin and other tissues.

Paper Details

Date Published: 6 June 2012
PDF: 6 pages
J. Biomed. Opt. 17(6) 066018 doi: 10.1117/1.JBO.17.6.066018
Published in: Journal of Biomedical Optics Volume 17, Issue 6
Show Author Affiliations
Daniel S. Gareau, Oregon Health Science Univ. (United States)
Anna Bar, Oregon Health & Science Univ. (United States)
Nicholas Snavely, Oregon Health & Science Univ. (United States)
Kenneth K. Lee, Oregon Health & Science Univ. (United States)
Neil Swanson, Oregon Health & Science Univ. (United States)
Eric Simpson, Oregon Health & Science Univ. (United States)
Steve Jacques, Oregon Health & Science Univ. (United States)
Nathaniel W. Chen, Oregon Health & Science Univ. (United States)


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