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Background/purposes Fluorescence emission from in vivo cutaneous melanin was recently detected under near-infrared ( NIR) excitation by our group. We then built a prototype NIR autofluorescence imaging system to observe and characterize the melanin distribution in human skin. In this article, we reported a new setup of NIR fluorescence imaging system and calibration methods to optimize the system for better clinical feasibility and clearer image. Methods The imaging system was designed to perform both fluorescence and reflectance imaging with a 785-nm fiber-coupled laser source. The illumination light was purified by a 785-nm bandpass filter for fluorescence excitation; while the spontaneous components were selected by a longpass filter for NIR reflectance imaging. A hand-controlled filter wheel was used to switch these two filters for different imaging modes. A dichroic filter was used to guide the illuminating light onto the skin surface for excitation. Reflectance and fluorescence signals were collected sequentially by a NIR optimized CCD camera. The captured images were calibrated by the reflectance images of a standard reflectance disk for non-uniform illuminations and light collection efficiencies. Results The clinical results demonstrated that NIR fluorescence intensities and distribution patterns vary among lesion types. It was also confirmed that pigmented skin lesions emitted higher NIR fluorescence than the surrounding normal skin due to the presentation of higher concentrations of cutaneous melanin within the lesions. Conclusion NIR autofluorescence imaging system could be utilized as a powerful tool for visualizing melanin distribution in pigmented skin lesions and as a potential method for aiding melanoma detection. [ABSTRACT FROM AUTHOR]

Abstract: Melanin content and distribution in skin were studied by examining a patient with white, brown and blue skin tones expressed on skin affected by vitiligo. Both diffuse reflectance and autofluorescence spectra of the three distinction skin sites were measured and compared. Monte Carlo simulations were then performed to help explain the measured spectral differences. The modeling is based on a six-layer skin optical model established from published skin optical parameters and by adding melanin content into different locations in the model skin. Both the reflectance and fluorescence spectra calculated by Monte Carlo (MC) simulation were approximately in agreement with experimental results. The study suggests that: (1) trichrome vitiligo skin may be an ideal in vivo model for studying the effect of skin melanin content and distribution on skin spectroscopy properties. (2) Based on the skin optical model and MC simulation, the content and distribution of melanin in skin, or other component of skin could be simulated and predicted. (3) Both reflectance and fluorescence spectra provided information about superficial skin structures but fluorescence spectra are capable of providing information from deeper cutaneous structures. (4) The research method, including the spectral ratio method, the method of adding and modifying the melanin content in skin optical models, and MC simulation could be applied in other non-invasive optical studies of the skin. [Copyright &y& Elsevier]

Objective measurements of melanin can provide important information for differentiating melanoma from benign pigmented lesions and in assessing pigmentary diseases. Herein, we evaluate near-infrared (NIR) fluorescence as a possible tool to quantify melanin. Various concentrations of in vitro Sepia melanin in tissue phantoms were measured with NIR fluorescence and diffuse reflectance spectroscopy. Similar optic measurements were conducted in vivo on 161 normal human skin sites. Diffuse reflectance spectroscopy was used to quantify the melanin content via Stamatas-Kollias algorithm. At physiologic concentrations, increasing in vitro melanin concentrations demonstrated higher fluorescence that was linearly correlated (R² = 0.99, p < .001). At higher concentrations, the fluorescence signal plateaued. A linear relationship was also observed with melanin content in human skin (R² = 0.59, p < .001). Comparing the fluorescence and reflectance signals with in vitro and in vivo samples, the estimated melanin concentration in human skin ranged between 0 and 1.25 mg/ml, consistent with previous quantitative studies involving invasive methods. [ABSTRACT FROM AUTHOR]

Background: Spectroscopic methods have been used to analyze in vivo melanin in the past but the specific effect of melanin depth on autofluorescence and reflectance spectroscopy has not been determined. In patients with blue vitiligo, three distinctive clinicopathologic patterns are present: (1) normal skin with normal epidermal melanin pigmentation (2), skin of blue vitiligo with dermal melanin pigmentation, and (3) tissue of regular vitiligo with no melanin pigmentation. Blue vitiligo may thus serve as an in vivo model to assess dermal pigment using spectroscopic techniques. Objectives: To evaluate the reflectance and autofluorescence spectra of a patient with blue vitiligo in order to assess the effect of melanin pigmentation and its localization on the optical properties of the skin. Methods: The blue-gray, normal and depigmented lesions of a patient with blue vitiligo were analyzed using reflectance and fluorescent spectroscopy. The condition was likely induced by a phototoxic reaction in a patient with pre-existing vitiligo. These data were then correlated to the histologic and electron microscopic findings present in the various types of lesions. Results: Reflectance spectroscopy detected little difference in spectral shape between skin sites affected by blue vitiligo vs. vitiligo. Autofluorescence spectroscopy detected an apparent difference between the two types of lesions, with the blue–gray lesions (blue vitiligo) showing lower fluorescence intensity and spectral maximum position red-shifted compared with regular vitiligo, whereas regular vitilgo showed more intense hemoglobin absorption than the blue vitiligo. Conclusions: Dermal melanin present in blue vitiligo can be well characterized by autofluorescence spectroscopy, while little difference in reflectance spectral shape exists between vitiligo and blue vitiligo. Thus, autofluorescence spectroscopy may better identify deeper structures in skin tissue, such as melanin, than reflectance spectroscopy [ABSTRACT FROM AUTHOR]