High-resolution functional imaging of the retina

项目摘要 为了了解活着的眼睛的视觉功能和细胞代谢，我们需要一种可视化的手段 并以非侵入性的方式评估分子动力学。其中涉及的一些关键分子 在视觉周期和细胞能量产生本质上是荧光的，但在 活着的眼睛通过单光子激发，因为所需的波长范围不是 通过眼睛的光学传输的。双光子激发荧光检眼镜(TPEFO)可以 用近红外光激发这些原本无法接触到的荧光团。在我们之前的实验中 在活的猕猴眼睛中，我们使用了自适应光学扫描光检眼镜(AOSLO)来 无创性地成像视网膜中许多不同类型的细胞，包括神经节细胞。 (Sharma，Williams等人，2016)。此外，我们还测量了视觉循环功能，通过表征 来自光感受器的双光子激发荧光的时间进程(Sharma，Schwarz等人，2016)。 我们现在建议将TPEFO在活的啮齿动物和猕猴眼睛中的能力扩大到 测量几个重要的荧光特性，如发射光谱、荧光寿命、 和氧化还原比率，这可能是细胞健康和功能的指标。这项研究有可能 提供对正常和改变的生化过程的洞察，并提高我们对 影响视网膜代谢和功能的疾病，如青光眼、黄斑变性和 Leber遗传性视神经病变。

Project Summary To understand visual function and cellular metabolism in the living eye, we need a means to visualize individual cells and assess molecular dynamics non-invasively. Some of the key molecules involved in the visual cycle and cellular energy production are intrinsically fluorescent, but are inaccessible in the living eye through single-photon excitation because the needed wavelength range is not transmitted by the optics of the eye. Two-photon excited fluorescence ophthalmoscopy (TPEFO) can excite these otherwise inaccessible fluorophores with near-infrared light. In our previous experiments in the living macaque eye, we used an adaptive optics scanning light ophthalmoscope (AOSLO) to non-invasively image many different cell classes throughout the retina, including ganglion cells (Sharma, Williams et al., 2016). Additionally, we measured visual cycle function by characterizing the time course of two-photon excited fluorescence from photoreceptors (Sharma, Schwarz et al., 2016). We now propose to expand the capabilities of TPEFO in the living rodent and macaque eye to measure several important fluorescence properties, such as emission spectrum, fluorescence lifetime, and redox ratio, which may be indicators of cell health and function. This research has the potential to provide insight into normal and altered biochemical processes and improve our understanding of diseases that impact retinal metabolism and function such as glaucoma, macular degeneration and Leber hereditary optic neuropathy.