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Journal of Biomedical Optics

Comparing <italic<in vivo</italic< pump–probe and multiphoton fluorescence microscopy of melanoma and pigmented lesions
Author(s): Jesse W. Wilson; Simone Degan; Christina S. Gainey; Tanya Mitropoulos; Mary Jane Simpson; Jennifer Y. Zhang; Warren S. Warren
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Paper Abstract

We demonstrate a multimodal approach that combines a pump–probe with confocal reflectance and multiphoton autofluorescence microscopy. Pump–probe microscopy has been proven to be of great value in analyzing thin tissue sections of pigmented lesions, as it produces molecular contrast which is inaccessible by other means. However, the higher optical intensity required to overcome scattering in thick tissue leads to higher-order nonlinearities in the optical response of melanin (e.g., two-photon pump and one-photon probe) that present additional challenges for interpreting the data. We show that analysis of pigment composition in vivo must carefully account for signal terms that are nonlinear with respect to the pump and probe intensities. We find that pump–probe imaging gives useful contrast for pigmented structures over a large range of spatial scales (100  μm to 1 cm), making it a potentially useful tool for tracking the progression of pigmented lesions without the need to introduce exogenous contrast agents.

Paper Details

Date Published: 21 November 2014
PDF: 13 pages
J. Biomed. Opt. 20(5) 051012 doi: 10.1117/1.JBO.20.5.051012
Published in: Journal of Biomedical Optics Volume 20, Issue 5
Show Author Affiliations
Jesse W. Wilson, Duke Univ. (United States)
Simone Degan, Duke Univ. (United States)
Duke Univ. Medical Ctr. (United States)
Christina S. Gainey, Duke Univ. (United States)
Tanya Mitropoulos, Duke Univ. (United States)
Mary Jane Simpson, Duke Univ. (United States)
Jennifer Y. Zhang, Duke Univ. (United States)
Warren S. Warren, Duke Univ. (United States)
Duke Univ. Medical Center (United States)


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