Special Section on Laser Applications in Life Sciences

Comparing in vivo pump–probe and multiphoton fluorescence microscopy of melanoma and pigmented lesions

[+] Author Affiliations
Jesse W. Wilson, Christina S. Gainey, Tanya Mitropoulos, Mary Jane Simpson

Duke University, Department of Chemistry, Box 90354, Durham, North Carolina 27708-0354, United States

Simone Degan

Duke University, Department of Chemistry, Box 90354, Durham, North Carolina 27708-0354, United States

Duke University Medical Center, Department of Radiology, Box 3808, Durham, North Carolina 27710, United States

Jennifer Y. Zhang

Duke University Medical Center, Department of Dermatology, DUMC 3135, Durham, North Carolina 27710, United States

Warren S. Warren

Duke University, Department of Chemistry, Box 90354, Durham, North Carolina 27708-0354, United States

Duke University Medical Center, Department of Radiology, Box 3808, Durham, North Carolina 27710, United States

Duke University, Department of Biomedical Engineering, Box 90281, Durham, North Carolina 27708, United States

J. Biomed. Opt. 20(5), 051012 (Nov 21, 2014). doi:10.1117/1.JBO.20.5.051012
History: Received July 5, 2014; Accepted October 30, 2014
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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 (100μm to 1 cm), making it a potentially useful tool for tracking the progression of pigmented lesions without the need to introduce exogenous contrast agents.

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© 2015 Society of Photo-Optical Instrumentation Engineers

Citation

Jesse W. Wilson ; Simone Degan ; Christina S. Gainey ; Tanya Mitropoulos ; Mary Jane Simpson, et al.
"Comparing in vivo pump–probe and multiphoton fluorescence microscopy of melanoma and pigmented lesions", J. Biomed. Opt. 20(5), 051012 (Nov 21, 2014). ; http://dx.doi.org/10.1117/1.JBO.20.5.051012


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