Fluorescence confocal mosaicing microscopy of basal cell carcinomas ex vivo: demonstration of rapid surgical pathology with high sensitivity and specificity

Daniel S. Gareau; Julie K. Karen; Stephen W. Dusza; Marie Tudisco; Kishwer S. Nehal; Milind Rajadhyaksha

doi:10.1117/12.809445

24 February 2009 Fluorescence confocal mosaicing microscopy of basal cell carcinomas ex vivo: demonstration of rapid surgical pathology with high sensitivity and specificity

Daniel S. Gareau, Julie K. Karen, Stephen W. Dusza, Marie Tudisco, Kishwer S. Nehal, Milind Rajadhyaksha

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Proceedings Volume 7169, Advanced Biomedical and Clinical Diagnostic Systems VII; 71690Y (2009) https://doi.org/10.1117/12.809445
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

Mohs surgery, for the precise removal of basal cell carcinomas (BCCs), consists of a series of excisions guided by the surgeon's examination of the frozen histology of the previous excision. The histology reveals atypical nuclear morphology, identifying cancer. The preparation of frozen histology is accurate but labor-intensive and slow. Nuclear pathology can be achieved by staining with acridine orange (1 mM, 20 s) BCCs in Mohs surgical skin excisions within 5-9 minutes, compared to 20-45 for frozen histology. For clinical utility, images must have high contrast and high resolution. We report tumor contrast of 10-100 fold over the background dermis and submicron (diffraction limited) resolution over a cm field of view. BCCs were detected with an overall sensitivity of 96.6%, specificity of 89.2%, positive predictive value of 93.0% and negative predictive value of 94.7%. The technique was therefore accurate for normal tissue as well as tumor. We conclude that fluorescence confocal mosaicing serves as a sensitive and rapid pathological tool. Beyond Mohs surgery, this technology may be extended to suit other pathological needs with the development of new contrast agents. The technique reported here accurately detects all subtypes of BCC in skin excisions, including the large nodular, small micronodular, and tiny sclerodermaform tumors. However, this technique may be applicable to imaging tissue that is larger, more irregular and of various mechanical compliances with further engineering of the tissue mounting and staging mechanisms.

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Daniel S. Gareau, Julie K. Karen, Stephen W. Dusza, Marie Tudisco, Kishwer S. Nehal, and Milind Rajadhyaksha "Fluorescence confocal mosaicing microscopy of basal cell carcinomas ex vivo: demonstration of rapid surgical pathology with high sensitivity and specificity", Proc. SPIE 7169, Advanced Biomedical and Clinical Diagnostic Systems VII, 71690Y (24 February 2009); https://doi.org/10.1117/12.809445

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KEYWORDS

Confocal microscopy

Tumors

Mohs surgery

Luminescence

Tissues

Skin

Surgery

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