High-Resolution Functional Imaging of the Retina

描述(申请人提供)：我们希望了解维持人类视觉的两个重要过程：视觉周期负责因吸收可见光而漂白的光色素的再生，以及每个活细胞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测量活着的眼睛中的分子动力学。其中一些分子本质上是荧光的，但用单光子荧光成像技术无法在活着的眼睛中获得，因为激发落在可以穿透眼睛光学的辐射范围之外。通过使用更长的激发波长，双光子激发荧光成像有可能激发这些原本无法获得的荧光团，并为成像许多视网膜结构提供固有的对比度。在我们的初步实验中，我们使用自适应光学扫描光检眼镜(AOSLO)从活的猕猴眼睛的锥体内节段成像双光子荧光(Hunter等人，2011年)。通过纠正眼睛的纵向色差和材料色散，我们计划提高我们成像系统的效率，并开发出可靠地成像眼睛多个视网膜层的结构和功能的方法。这一能力不仅将提供对正常细胞马赛克及其生化过程的洞察，还将提高我们对许多影响这些视网膜生化级联反应的疾病的了解，例如Stargardt病、黄斑变性和Leber遗传性视神经病变。 与公共健康相关：我在罗切斯特大学的新实验室将把双光子荧光成像开发成一种高效的技术，首次以非侵入性的方式显示眼后的神经细胞，并监测活着的眼睛中重要的细胞过程。这项研究可能会改进一些视网膜疾病的诊断，监测其进展情况，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.