In vivo imaging with a fast large-area multiphoton exoscope (FLAME) captures the melanin distribution heterogeneity in human skin.
In vivo imaging with a fast large-area multiphoton exoscope (FLAME) captures the melanin distribution heterogeneity in human skin.
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DOI：
10.1038/s41598-022-12317-y
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发表时间：
2022-05-16
影响因子：
4.6
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中科院分区：
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黑色素在调节表皮稳态和皮肤光保护中起着重要作用。由于其参与广泛的生理和病理皮肤过程，对其表皮分布和总体含量的评估引起了人们的极大兴趣。在已经报道的用于人类皮肤中黑色素的非侵入性定量的几种光谱和光学成像方法中，基于双光子激发的荧光寿命的检测的方法通过使得能够以亚细胞分辨率选择性地检测黑色素来区分自身，从而促进其定量，同时还解析其深度分布。基于这种方法的黑色素评估的现有研究的关键限制是它们不能解释由于图像的视场减小而导致的皮肤异质性，这导致测量值的高度分散。正常和病理人类皮肤中的色素沉着是高度异质性的，并且其宏观定量对于表皮黑色素分布的可靠测量和捕获作为对治疗的响应的黑色素相关的敏感动态变化是至关重要的。在这项工作中，我们采用了快速的大面积多光子外镜（FLAME），最近开发的临床皮肤成像，有能力评估的3D分布的表皮黑色素含量在体内宏观（毫米尺度）与微观分辨率（亚微米）和快速采集速率（分钟）。我们证明了通过捕获大体积采样实现的受试者内色素沉着异质性，在菲茨帕特里克I至V型皮肤中的黑色素密度和分布测量的可靠性显著增强。我们还证明了这种方法的潜力，以提供一致的测量结果时，在不同的时间成像相同的皮肤区域。这些进展对于与监测色素调节相关的临床和研究应用至关重要，作为对色素性皮肤病，皮肤老化以及皮肤癌治疗的反应。
Melanin plays a significant role in the regulation of epidermal homeostasis and photoprotection of human skin. The assessment of its epidermal distribution and overall content is of great interest due to its involvement in a wide range of physiological and pathological skin processes. Among several spectroscopic and optical imaging methods that have been reported for non-invasive quantification of melanin in human skin, the approach based on the detection of two-photon excited fluorescence lifetime distinguishes itself by enabling selective detection of melanin with sub-cellular resolution, thus facilitating its quantification while also resolving its depth-profile. A key limitation of prior studies on the melanin assessment based on this approach is their inability to account for the skin heterogeneity due to the reduced field of view of the images, which results in high dispersion of the measurement values. Pigmentation in both normal and pathological human skin is highly heterogeneous and its macroscopic quantification is critical for reliable measurements of the epidermal melanin distribution and for capturing melanin-related sensitive dynamic changes as a response to treatment. In this work, we employ a fast large-area multiphoton exoscope (FLAME), recently developed by our group for clinical skin imaging, that has the ability to evaluate the 3D distribution of epidermal melanin content in vivo macroscopically (millimeter scale) with microscopic resolution (sub-micron) and rapid acquisition rates (minutes). We demonstrate significant enhancement in the reliability of the melanin density and distribution measurements across Fitzpatrick skin types I to V by capturing the intra-subject pigmentation heterogeneity enabled by the large volumetric sampling. We also demonstrate the potential of this approach to provide consistent measurement results when imaging the same skin area at different times. These advances are critical for clinical and research applications related to monitoring pigment modulation as a response to therapies against pigmentary skin disorders, skin aging, as well as skin cancers.
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