Super-resolution ophthalmoscopy for in vivo retinal imaging

用于体内视网膜成像的超分辨率检眼镜

• 批准号: 8975948
• 负责人: XINCHENG YAO
• 金额: $ 29.23万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8975948
• 关键词: Super; resolution; ophthalmoscopy; vivo; retinal

DESCRIPTION (provided by applicant): This 3-year R01 project is to explore a new methodology for super-resolution scanning laser ophthalmoscopy (SLO). We have demonstrated resolution-doubling in scanning laser microscopy (SLM) and optical coherence tomography (OCT) through virtually structured detection (VSD). We propose here to extend the VSD into super-resolution ophthalmoscopy for in vivo retinal imaging. Without the complexity of structured illumination microscopy (SIM), the VSD provides an easy, low-cost and phase-artifact free strategy to achieve super- resolution imaging. However, deployable application of the VSD for in vivo retinal imaging is challenged by limited frame-speed (40s per frame) of our prototype instrument. Such low speed is intolerable for in vivo retinal imaging because of within-frame blur due to eye movements. We propose here to combine rapid line-scan strategy and accurate image registration to compensate for eye movements. The first aim of this project is to construct a line-scan super-resolution SLO. Success criterion of this aim is to produce an instrument that enables super-resolution imaging at 10 ms frame-speed. In order to achieve rapid change of three-angle (0o, 30o and 60o) scanning required for super-resolution reconstruction, a galvo-dove-prism will be assembled. Preliminary line-scan imaging of frog eyes indicates that within-frame blur can be ignored at 10 ms frame- speed; while frame-by-frame movement can be corrected by accurate image registration. The second aim is to conduct both ex vivo and in vivo validation of the proposed instrument. Before in vivo experiments, a 10x objective with numeric aperture NA=0.25 will be used for super-resolution microscopy of 20 nm nanoparticles. The ex vivo experiment is designed to verify spatial resolution of the super-resolution instrument. The second aim of this project is to validate the super-resolution SLO for in vivo retinal imaging of anesthetized animals (frogs). In vivo imaging of anesthetized frog will be conducted to quantify resolution difference in super- resolution ophthalmoscopy and conventional SLO. The third aim to verify transient rod phototropism (TRP) in intact animals. We have recently demonstrated TRP in freshly isolated mouse and frog retinas. In vivo verification of the TRP will provide an optical biomarker suitable for functional mapping of rod physiology at cellular resolution. Successful endpoint of this project is to demonstrate the feasibility of super-resolution ophthalmoscopy of intact animals. In future phase of this project, the super-resolution ophthalmoscopy will be used to investigate biophysical mechanism of the TRP in normal animals, and to characterize abnormal modification of the TRP in transgenic animals with photoreceptor degeneration. Moreover, we also plan to combine adaptive optics with the proposed instrument to pursue super-resolution ophthalmoscopy of human subjects in following phase of this project.

描述（由申请人提供）：本3年R 01项目旨在探索超分辨率扫描激光检眼镜（SLO）的新方法。我们已经证明了分辨率加倍扫描激光显微镜（SLM）和光学相干断层扫描（OCT）通过虚拟结构检测（VSD）。我们建议在这里扩展VSD到超分辨率检眼镜在体内视网膜成像。没有结构照明显微镜（SIM）的复杂性，VSD提供了一种简单、低成本且无相位伪影的策略来实现超分辨率成像。然而，VSD用于体内视网膜成像的可部署应用受到我们的原型仪器的有限帧速度（每帧40秒）的挑战。由于眼睛运动引起的帧内模糊，这种低速度对于体内视网膜成像是不可容忍的。我们在这里建议结合联合收割机快速线扫描策略和准确的图像配准，以补偿眼球运动。该项目的首要目标是构建线扫描超分辨率SLO。这一目标的成功标准是生产一种能够以10 ms帧速度进行超分辨率成像的仪器。为了实现超分辨率重建所需的三个角度（0 °、30 °和60 °）扫描的快速变化，将组装一个振镜-鸽子-棱镜。蛙眼的初步线扫描成像表明，在10 ms的帧速度下，帧内模糊可以被忽略;而逐帧运动可以通过精确的图像配准来校正。第二个目的是对所提出的仪器进行体外和体内验证。在体内实验之前，数值孔径NA=0.25的10倍物镜将用于20 nm纳米颗粒的超分辨率显微镜检查。设计了离体实验来验证超分辨率仪器的空间分辨率。该项目的第二个目的是验证用于麻醉动物（青蛙）的活体视网膜成像的超分辨率SLO。将对麻醉青蛙进行体内成像，以量化超分辨率检眼镜和传统SLO的分辨率差异。第三个目的是在完整动物中验证瞬时视杆细胞向光性（TRP）。我们最近在新鲜分离的小鼠和青蛙视网膜中证明了TRP。TRP的体内验证将提供适合于在细胞分辨率下进行视杆生理学功能映射的光学生物标志物。该项目的成功终点是证明在完整动物中进行超分辨率检眼镜检查的可行性。在本项目的未来阶段，超分辨率检眼镜将用于研究正常动物中TRP的生物物理机制，并表征感光细胞变性的转基因动物中TRP的异常修饰。此外，我们也计划在本计画的后续阶段中，将联合收割机自适应光学与所提出的仪器结合，以追求人类受试者的超解析检眼镜。