High-Resolution Functional Imaging of the Retina

项目摘要 我们希望了解维持人类视觉的两个重要过程;视觉周期是 负责通过吸收可见光和细胞色素而漂白的色素的再生 每个活细胞都需要新陈代谢来提供能量。为了研究这些过程，我们需要一个 方法来可视化单个细胞和测量分子动力学在活的眼睛。一些 所涉及的分子本质上是荧光的，但在活体眼睛中是无法用单- 光子荧光成像，因为激发福尔斯落在可以 穿透眼睛的光学系统。通过使用相当长的激发波长， 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固有对比度。在最初的实验中，我们 使用自适应光学扫描光检眼镜（AOSLO）对双光子荧光成像 来自活猕猴眼睛中的视锥内节（Hunter等人，2011）。通过校正 纵向色差和材料色散的眼睛，我们计划提高效率 我们的成像系统和开发方法，可靠地成像的结构和功能， 眼睛里的多层视网膜。这种能力不仅可以提供对正常细胞镶嵌的深入了解， 以及它们的生化过程，它也将提高我们对许多疾病的理解， 这些视网膜生化级联反应，如Stargardt病，黄斑变性和Leber病， 遗传性视神经病变

Project Summary 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 into 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.