High-resolution functional imaging of the retina

项目摘要 为了了解活体眼睛的视觉功能和细胞代谢，我们需要一种可视化的方法， 单个细胞并非侵入性地评估分子动力学。一些关键的分子 在视觉周期和细胞能量生产本质上是荧光的，但在 因为所需的波长范围不 通过眼睛的光学系统传输。双光子激发荧光检眼镜检查（TPEFO）可以 用近红外光激发这些原本难以接近的荧光团。在我们之前的实验中 在活体猕猴眼中，我们使用自适应光学扫描光检眼镜（AOSLO）， 非侵入性成像许多不同的细胞类整个视网膜，包括神经节细胞 （Sharma，威廉姆斯等人，2016）。此外，我们还通过表征 来自光感受器的双光子激发荧光的时间过程（Sharma，施瓦茨等人，2016）。 我们现在建议扩大TPEFO在活体啮齿动物和猕猴眼睛中的能力， 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，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物化学过程的深入了解，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.