小鼠视网膜的无标记非线性光学成像
Label-free nonlinear optical imaging of mouse retina.
作者信息
He Sicong, Ye Cong, Sun Qiqi, Leung Christopher K S, Qu Jianan Y
机构信息
Department of Electronic and Computer Engineering, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China ; Center of Systems Biology and Human Health, School of Science and Institute for Advanced Study, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong SAR, China ; These authors contributed equally to this work.
Department of Ophthalmology & Visual Sciences, The Chinese University of Hong Kong, Kowloon, Hong Kong, China ; These authors contributed equally to this work.
出版信息
Biomed Opt Express. 2015 Feb 26;6(3):1055-66. doi: 10.1364/BOE.6.001055. eCollection 2015 Mar 1.
A nonlinear optical (NLO) microscopy system integrating stimulated Raman scattering (SRS), two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG) was developed to image fresh mouse retinas. The morphological and functional details of various retinal layers were revealed by the endogenous NLO signals. Particularly, high resolution label-free imaging of retinal neurons and nerve fibers in the ganglion cell and nerve fiber layers was achieved by capturing endogenous SRS and TPEF signals. In addition, the spectral and temporal analysis of TPEF images allowed visualization of different fluorescent components in the retinal pigment epithelium (RPE). Fluorophores with short TPEF lifetime, such as A2E, can be differentiated from other long-lifetime components in the RPE. The NLO imaging method would provide important information for investigation of retinal ganglion cell degeneration and holds the potential to study the biochemical processes of visual cycle in the RPE.
开发了一种集成受激拉曼散射（SRS）、双光子激发荧光（TPEF）和二次谐波产生（SHG）的非线性光学（NLO）显微镜系统，用于对新鲜小鼠视网膜成像。内源性NLO信号揭示了各视网膜层的形态和功能细节。特别地，通过捕获内源性SRS和TPEF信号，实现了对神经节细胞层和神经纤维层中视网膜神经元和神经纤维的高分辨率无标记成像。此外，TPEF图像的光谱和时间分析使得视网膜色素上皮（RPE）中不同荧光成分得以可视化。具有短TPEF寿命的荧光团，如A2E，可以与RPE中其他长寿命成分区分开来。NLO成像方法将为研究视网膜神经节细胞变性提供重要信息，并具有研究RPE中视觉循环生化过程的潜力。
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