High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution
High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution
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DOI：
10.1117/1.1577349
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发表时间：
2003-07-01
影响因子：
3.5
通讯作者：
Riemann, I
中科院分区：
文献类型：
--
作者：
König, K;Riemann, I
基于多光子自发荧光成像和二次谐波产生（SHG）的人体皮肤高分辨率四维（4-D）光学层析成像与紧凑型飞秒激光成像系统DermaInspect以及改进的多光子显微镜进行。飞秒激光脉冲的80 MHz的光谱范围为750至850 nm，快速galvoscan镜，和时间相关的单光子计数模块已被用来成像人体皮肤在体外和体内亚细胞的空间和250 ps的时间分辨率。非线性诱导的自发荧光来源于天然内源性荧光团和蛋白质结构，例如还原的烟酰胺腺嘌呤二核苷酸磷酸、黄素、胶原蛋白、弹性蛋白、卟啉和黑色素。二次谐波产生用于检测胶原结构。已经研究了患有皮肤病如银屑病、真菌感染、痣和黑色素瘤的患者的组织。单个组织内细胞和皮肤结构可以清楚地可视化。细胞内组分和结缔组织结构可以进一步通过荧光激发光谱、通过测定每个像素的荧光衰减和通过荧光寿命成像来表征。新型非侵入性多光子自体荧光SHG成像技术提供了具有亚细胞分辨率的4-D（x，y，z，tau）光学活检，并提供了在皮肤病学中引入高分辨率光学诊断方法的可能性。(C)2003年，由光学仪器工程师学会（Society of Photo-Optical Instrumentation Engineers）主办。
High-resolution four-dimensional (4-D) optical tomography of human skin based on multiphoton autofluorescence imaging and second harmonic generation (SHG) was performed with the compact femtosecond laser imaging system DermaInspect as well as a modified multiphoton microscope. Femtosecond laser pulses of 80 MHz in the spectral range of 750 to 850 nm, fast galvoscan mirrors, and a time-correlated single-photon counting module have been used to image human skin in vitro and in vivo with subcellular spatial and 250-ps temporal resolution. The nonlinear induced autofluorescence originates from naturally endogenous fluorophores and protein structures such as reduced nicotinamide adenine dinucleotide phosphate, flavins, collagen, elastin, porphyrins, and melanin. Second harmonic generation was used to detect collagen structures. Tissues of patients with dermatological disorders such as psoriasis, fungal infections, nevi, and melanomas have been investigated. Individual intratissue cells and skin structures could be clearly visualized. Intracellular components and connective tissue structures could be further characterized by fluorescence excitation spectra, by determination of the fluorescence decay per pixel, and by fluorescence lifetime imaging. The novel noninvasive multiphoton autofluorescence-SHG imaging technique provides 4-D (x,y,z,tau) optical biopsies with subcellular resolution and offers the possibility of introducing a high-resolution optical diagnostic method in dermatology. (C) 2003 Society of Photo-Optical Instrumentation Engineers.