Interferometric Optophysiology of the Human Retina

项目摘要/摘要 我们的目标是开发一种非侵入性光学监测个体视网膜活动的新技术 活着的人类视网膜中的神经元和它们的光驱动输入，在细胞分辨率上。如果成功，这将是 技术将提供一种全新的、客观的方法来了解和监测视网膜的治疗 疾病，从而改变了对眼睛和视力的科学研究。该项目直接解决了 在RFA-EY-14-001中概述的优先事项，这是NEI大胆目标倡议范围内的第一个RFA。 拟议的工作依赖于结合和验证两种新方法。第一，干涉测量 (包括相分辨OCT；加州大学河滨分校的Park Lab)原则上可以用于测量纳米- 在细胞膜去极化和离子内流过程中发生的细胞膜鳞片扭曲。使用 深度分辨率，这些测量将使我们能够非侵入性地测量整个 视网膜的各层，在细胞分辨率下。第二，自适应光学扫描激光眼底镜(Roorda Lab 在加州大学伯克利分校)和基于图像的眼睛跟踪可以用于定位刺激和测量光束 通过克服光学像差和眼睛抖动，在活体眼睛中具有细胞精确度的视网膜。这 技术将允许我们用可见光激活单个光感受器和一组光感受器 同时成像单个下游细胞在体内产生的电活动。 要推进和结合这些方法，需要逐步聚合技术。在一个 独特的合作，我们将建立在更简单的广域干涉测量电活动的基础上 分离视网膜(斯坦福大学Palanker实验室)，结合大规模多电极生理学 在灵长类动物视网膜中进行测量(斯坦福大学奇奇尔尼斯基实验室)以验证和调整光学 测量。 最终，每一步的创新都形成了一个强大的工具，无论是独立的还是与以下各项相结合的 其他方法，并发现适用于光学成像、视网膜生理学、心理物理学和临床 眼科。具体目标是： 目的1.测量灵长类动物视网膜电活动模式的广场干涉法 神经信号传递过程中的去极化会在细胞中产生纳米级的变形，这些变形可以通过 干涉计量学。最简单的方法是具有传输几何结构的广域干涉显微镜。 孤立的视网膜。我们将测量灵长类动物神经活动产生的深度分辨光学相变 视网膜，并用于许多视网膜神经节细胞(RGC)和其他视网膜的生理特性 神经元同时存在。 目的2.相位分辨OCT用于视网膜活动模式的反射测量 人类应用的下一步是相位分辨OCT；本质上，低相干干涉测量和 用于活体成像的成熟工具。我们将记录与神经相关的光路长度变化 使用点扫描、近红外(1060 Nm)、相分辨OCT在孤立的OCT上的反射几何活动 灵长类视网膜。 目的3.用于测量人类视网膜功能的自适应光学、眼球跟踪和相位分辨OCT 在人类身上部署需要补偿眼睛的光学像差和眼睛运动。 我们将开发一种系统，使用AOSLO对视网膜进行成像，以进行眼睛跟踪，有针对性地传递刺激 光，以及OCT探头的定位。我们将在人体上测试这个系统，并展示它的潜力 在临床环境中的应用。

Project Summary/Abstract Our goal is to develop a new technology for non-invasive optical monitoring of activity of individual retinal neurons and their light-driven inputs, at cellular resolution, in the living human retina. If successful, this technology will provide an entirely new and objective approach to understand and monitor treatment of retinal disease, thereby transforming scientific studies of the eye and vision. This project directly addresses the priorities outlined in the RFA-EY-14-001, the first RFA within the NEI Audacious Goal Initiative. The proposed work relies on combining and validating two new approaches. First, interferometry (including phase-resolved OCT; Park Lab at UC Riverside) can, in principle, be used to measure nanometer- scale distortions in the membranes of cells that occur during membrane depolarization and ion influx. With depth resolution, these measurements will enable us to measure neural activity non-invasively, throughout the layers of the retina, at cellular resolution. Second, adaptive optics scanning laser ophthalmoscopy (Roorda Lab at UC Berkeley) and image-based eye tracking can be used to position stimulating and measurement beams on the retina with cellular precision in the living eye, by overcoming optical aberrations and eye jitter. This technology will allow us to activate individual photoreceptors and groups of photoreceptors with visible light while imaging the resulting electrical activity of individual downstream cells, in vivo. To advance and combine these approaches requires a stepwise aggregation of technology. In a unique collaboration, we will build on simpler wide-field interferometric measurements of electrical activity in isolated retina (Palanker Lab at Stanford University), combined with large-scale multi-electrode physiological measurements in primate retina (Chichilnisky Lab at Stanford University) to validate and tune the optical measurements. Ultimately, the innovation at each step forms a powerful tool, independently or with a combination of other approaches, and finds applicability to optical imaging, retinal physiology, psychophysics and clinical ophthalmology. The specific aims are: Aim 1. Wide-field interferometry for measuring patterns of electrical activity in primate retina Depolarization during neural signaling produces nanometer-scale deformations in cells that are detectable with interferometry. The simplest approach is wide-field interferometric microscopy with transmission geometry in isolated retina. We will measure depth-resolved optical phase changes produced by neural activity in primate retina, and use them for physiological characterizations of many retinal ganglion cells (RGCs) and other retinal neurons simultaneously. Aim 2. Phase-resolved OCT for reflectance measurements of patterns of retinal activity The next step toward human application is phase-resolved OCT; essentially, low-coherence interferometry and well-established tool for in vivo imaging. We will record optical path length changes associated with neural activity in reflection geometry using point-scanning, near-IR (1060 nm), phase-resolved OCT on isolated primate retina. Aim 3. Adaptive optics, eye tracking and phase-resolved OCT for measuring human retinal function Deployment in humans requires compensating for optical aberrations in the eye as well as eye movements. We will develop a system that uses AOSLO to image the retina for eye tracking, targeted delivery of stimulation light, and positioning of the OCT probe. We will test this system in humans and demonstrate its potential application in clinical settings.