High-Resolution Functional Imaging of the Retina

描述（由申请人提供）：我们希望了解维持人类视觉的两个重要过程;视觉周期负责通过吸收可见光而漂白的色素的再生，并且每个活细胞都需要细胞代谢来提供能量。为了研究这些过程，我们需要一种方法来可视化单个细胞并在活体眼睛中测量分子动力学。所涉及的一些分子本质上是荧光的，但是在活体眼睛中用单光子荧光成像是不可接近的，因为激发福尔斯落在可以穿透眼睛的光学器件的辐射范围之外。通过使用相当长的激发波长，双光子激发荧光成像具有激发这些否则难以接近的荧光团的潜力，并提供用于成像许多视网膜结构的内在对比度。在我们最初的实验中，我们使用自适应光学扫描光检眼镜（AOSLO）来对来自活猕猴眼睛中的视锥内节的双光子荧光进行成像（Hunter等人，2011）。通过校正眼睛的纵向色差和材料色散，我们计划提高成像系统的效率，并开发用于可靠地成像眼睛中多个视网膜层的结构和功能的方法。这种能力不仅将提供对正常细胞镶嵌及其生化过程的深入了解，还将提高我们对影响这些视网膜生化级联反应的许多疾病的理解，例如Stargardt病，黄斑变性和Leber遗传性视神经病变。 公共卫生相关性：我在罗切斯特大学的新实验室将开发双光子荧光成像技术，使其成为一种高效的技术，首次非侵入性地观察眼睛后部的神经细胞，并监测活体眼睛中的重要细胞过程。这项研究可能会改善一些视网膜疾病的诊断，监测其进展并建立治疗策略的有效性。

DESCRIPTION (provided by applicant): We wish to understand two important processes that sustain human vision; the visual cycle is responsible for regeneration of photopigment bleached by absorption of visible light and cellular metabolism is required in every living cell to provide energy. To study these processes, we need a method to visualize individual cells and measure molecular dynamics in the living eye. Some of the molecules involved are intrinsically fluorescent, but are inaccessible in the living eye with single- photon fluorescence imaging because the excitation falls outside the range of radiation that can penetrate the optics of the eye. By using considerably longer excitation wavelengths, two-photon excited fluorescence imaging has the potential to excite these otherwise inaccessible fluorophores and provide intrinsic contrast for imaging a number of retinal structures. In our initial experiments, we used an adaptive optics scanning light ophthalmoscope (AOSLO) to image two-photon fluorescence from cone inner segments in the living macaque eye (Hunter et al, 2011). By correcting the longitudinal chromatic aberration and material dispersion of the eye, we plan to improve the efficiency of our imaging systems and develop methodology for reliably imaging both structure and function of multiple retinal layers in the eye. Not only will this capability provide insight ito normal cell mosaics and their biochemical processes, it will also improve our understanding of many diseases that impact these retinal biochemical cascades such as Stargardt's disease, macular degeneration and Leber's hereditary optic neuropathy. PUBLIC HEALTH RELEVANCE: My new laboratory at the University of Rochester will develop two-photon fluorescence imaging into a highly efficient technique to visualize non-invasively for the first time neural cells in the back of the eye and to monitor important cellular processes in the living eye. This research could lead to improvements in the diagnosis of some retinal diseases, monitoring of their progression and establishing the efficacy of treatment strategies.