Journal
JOURNAL OF BIOMEDICAL OPTICS
Volume 20, Issue 5, Pages -Publisher
SPIE-SOC PHOTO-OPTICAL INSTRUMENTATION ENGINEERS
DOI: 10.1117/1.JBO.20.5.051012
Keywords
in vivo microscopy; melanoma; pump-probe microscopy; multiphoton microscopy; time-resolved spectroscopy; transient absorption
Funding
- NIH [1RC1CA145105, R01-CA166555, P30-AR066527, 1F32CA168497-01A1]
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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 mu m to 1 cm), making it a potentially useful tool for tracking the progression of pigmented lesions without the need to introduce exogenous contrast agents. (C) 2015 Society of Photo-Optical Instrumentation Engineers (SPIE)
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