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Multiphoton microscopy (MPM) is a technology that can generate real-time, depth-resolved subsurface images of skin with histologic resolution and sensitivity based on endogenous molecular and chemical contrast. In skin, MPM contrast is derived from second harmonic generation (SHG) of collagen and two-photon excited fluorescence (TPEF) of autofluorescent co-factors NAD(P)H and FAD, elastin, keratin, and melanin. In addition to the specificity provided by the detection of the SHG signal from collagen, MPM can visualize specific skin fluorophores based on their fluorescence lifetime detection. Over the past several years, our group and others have demonstrated the MPM strong potential for a broad range of applications from advancing the understanding of skin biology to non-invasive diagnosis of skin diseases and monitoring therapy effects. However, a routine implementation of this technology in clinical research and practice requires advancing the instrumentation to allow for easier access by clinicians and for more efficient imaging in terms of speed and scanning area. Our group recently introduced benchtop innovations and developed a fast large area multiphoton exoscope (FLAME) that can provide rapid, real-time, depth-resolved images of skin, over macroscopic areas (cm-scale) with microscopic resolution (0.5-1m) and chemical contrast (selective detection of melanin). This presentation will highlight the latest advances in the FLAME development, including its conversion into a compact, portable device, highly optimized for clinical skin imaging with enhanced sensitivity and specificity. The technical abilities of this imaging platform will be demonstrated along with the results of a study that establishes clinical safety and imaging performance in 20 volunteers with normal skin. The results emphasize the significance of macroscopic imaging in the context of skin heterogeneity in terms of pigment distribution and dermal photodamage.
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