Confocal mosaicing microscopy of basal-cell carcinomas ex vivo: progress in digital staining to simulate histology-like appearance

Jason Bini; James Spain; Kishwer Nehal; Vikki Hazelwood; Charles DiMarzio; Milind Rajadhyaksha

doi:10.1117/12.873601

21 February 2011 Confocal mosaicing microscopy of basal-cell carcinomas ex vivo: progress in digital staining to simulate histology-like appearance

Jason Bini, James Spain, Kishwer Nehal, Vikki Hazelwood, Charles DiMarzio, Milind Rajadhyaksha

Author Affiliations +

Proceedings Volume 7890, Advanced Biomedical and Clinical Diagnostic Systems IX; 78900E (2011) https://doi.org/10.1117/12.873601
Event: SPIE BiOS, 2011, San Francisco, California, United States

Abstract

Confocal mosaicing microscopy enables rapid imaging of large areas of fresh tissue, without the processing that is necessary for conventional histology. Using acridine orange (1 milliMolar, 20 seconds) to stain nuclei, basal cell carcinomas were detected in fluorescence confocal mosaics of Mohs surgical excisions with sensitivity of 96.6% and specificity of 89.2%. A possible barrier toward clinical acceptance is that confocal mosaics are based on a single mode of contrast and appear in grayscale, whereas histology is based on two (hematoxylin for nuclei, eosin for cellular cytoplasm and dermis) and appears purple-and-pink. Toward addressing this barrier, we report progress in developing a multispectral analytical model for digital staining: fluorescence confocal mosaics, which show only nuclei, are digitally stained purple and overlaid on reflectance confocal mosaics, which show only cellular cytoplasm and dermis, and digitally stained pink, to mimic the appearance of histology. Comparison of digitally stained confocal mosaics by our Mohs surgeon to the corresponding Mohs histology shows good correlation for normal and tumor detail. Digitally stained confocal mosaicing microscopy may allow direct examination of freshly excised tissue and serve as an adjunct for rapid pathology at-the-bedside.

Citation Download Citation

Jason Bini, James Spain, Kishwer Nehal, Vikki Hazelwood, Charles DiMarzio, and Milind Rajadhyaksha "Confocal mosaicing microscopy of basal-cell carcinomas ex vivo: progress in digital staining to simulate histology-like appearance", Proc. SPIE 7890, Advanced Biomedical and Clinical Diagnostic Systems IX, 78900E (21 February 2011); https://doi.org/10.1117/12.873601

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