title: Comparing in vivo pump-probe and multiphoton fluorescence microscopy of melanoma and pigmented lesions.
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.
authors.affiliation: Duke University, Department of Chemistry, Box 90354, Durham, North Carolina 27708-0354, United States.
authors.forename: Jesse W
authors.initials: JW
authors.lastname: Wilson
authors.affiliation: Duke University, Department of Chemistry, Box 90354, Durham, North Carolina 27708-0354, United StatesbDuke University Medical Center, Department of Radiology, Box 3808, Durham, North Carolina 27710, United States.
authors.forename: Simone
authors.initials: S
authors.lastname: Degan
authors.affiliation: Duke University, Department of Chemistry, Box 90354, Durham, North Carolina 27708-0354, United States.
authors.forename: Christina S
authors.initials: CS
authors.lastname: Gainey
authors.affiliation: Duke University, Department of Chemistry, Box 90354, Durham, North Carolina 27708-0354, United States.
authors.forename: Tanya
authors.initials: T
authors.lastname: Mitropoulos
authors.affiliation: Duke University, Department of Chemistry, Box 90354, Durham, North Carolina 27708-0354, United States.
authors.forename: Mary Jane
authors.initials: MJ
authors.lastname: Simpson
authors.affiliation: Duke University Medical Center, Department of Dermatology, DUMC 3135, Durham, North Carolina 27710, United States.
authors.forename: Jennifer Y
authors.initials: JY
authors.lastname: Zhang
authors.affiliation: Duke University, Department of Chemistry, Box 90354, Durham, North Carolina 27708-0354, United StatesbDuke University Medical Center, Department of Radiology, Box 3808, Durham, North Carolina 27710, United StatesdDuke University, Department of Biomedical.
authors.forename: Warren S
authors.initials: WS
authors.lastname: Warren
mesh_terms: D000818:Animals; D003287:Contrast Media / Q000737:chemistry; D006801:Humans; D015603:Keratinocytes / Q000166:cytology; D008543:Melanins / Q000737:chemistry; D008544:Melanocytes / Q000166:cytology; D008545:Melanoma / Q000473:pathology*; D051379:Mice; D008853:Microscopy / Q000295:instrumentation / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D009368:Neoplasm Transplantation; D017785:Photons; D012867:Skin / Q000473:pathology; D012878:Skin Neoplasms / Q000473:pathology*; D012880:Skin Pigmentation / Q000502:physiology*
chemical_list: D003287:Contrast Media; D008543:Melanins