Non-invasive, living histology of capillary structure and single cell blood flow in mouse model of diabetic retinopathy

糖尿病视网膜病变小鼠模型毛细血管结构和单细胞血流的非侵入性活体组织学研究

• 批准号: 10213738
• 负责人: Jesse Barrett Schallek
• 金额: $ 40.42万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10213738
• 关键词: Adult; Age; Animals; Back; Blindness; Blood; Blood Cells; Blood Glucose; Blood Vessels; Blood capillaries; Blood flow; Caliber; Cells; Chronic; Clinical; Contrast Media; Coupled; Data; Detection; Diabetes Mellitus; Diabetic Retinopathy; Diabetic mouse; Disease; Event; Excision; Eye; Eye Manifestations; Eye diseases; Functional disorder; Hair; Histology; Human; Hyperglycemia; Image; Impairment; Individual; Insulin-Dependent Diabetes Mellitus; Legal patent; Light; Measures; Microscope; Microscopic; Morphology; Motion; Movement; Mus; Natural History; Ophthalmoscopes; Ophthalmoscopy; Optics; Outcome; Pathologic; Pathology; Pattern; Perfusion; Pericytes; Pharmacology; Population; Property; Public Health; Regional Perfusion; Research Project Grants; Resolution; Retina; Retinal Diseases; Scanning; Shunt Device; Speed; Stress; Structure; Testing; Thick; Time; Tissues; Transgenic Organisms; Vascular Diseases; Velocimetries; adaptive optics; arteriole; base; cell type; cellular targeting; design; diabetic; high resolution imaging; human model; innovation; light scattering; mouse model; new technology; non-invasive imaging; response; retina blood vessel structure; retina circulation; software development; venule; wasting

Summary/Abstract In the US, diabetic retinopathy is the leading cause of blindness in working age adults and remains a public health problem throughout the world. The earliest manifestations of this eye disease are believed to originate in capillary dysfunction resulting in both over- and under perfusion of regional capillaries. And while these changes in microvascular structure have been identified as hallmarks of the disease, the earliest functional changes in this microscopic network remain unclear. Is microvascular flow impaired early before capillary structural changes, or does the formation of aberrant vessel patterns, as a consequence, profoundly change retinal capillary flow? Conventional retina cameras generally lack the necessary resolution to study capillary-level blood flow because the eye's optics blur the microscopic capillaries at the back of the eye. In this study, we develop and deploy a new retinal camera that turns the eye into a high-power microscope to study single cell blood flow the back of the living eye. Combined with the optical improvements of this adaptive optics camera which corrects for image blur, we have coupled two other innovations to image the movement of individual blood cells as they move through the tiniest of capillaries only 1/10th the thickness of a human hair. First, blood cells are not only microscopic, but they also move at fast rates of speed. To image these blood cells free of motion blur, the use of a high-speed camera is required. In this research project, we combine the blur-correcting optics with an exceptionally fast camera that can capture over 30,000 snapshots per second. This camera is focused at single capillaries and can image the blood cells as they flow by -one by one. This advancement allows us to measure blood cell speed and provide exact counts of the number of passing blood cells, two innovative measures of blood flow at the capillary level. A second innovation uses special light-scattering properties of blood cells to provide highly detailed images of blood cell boundaries against the vessel wall and surrounding tissue. The resultant images provide not only high resolution images of blood cells, but can also provide unprecedented measures of blood cell type and their deformation within microvessels of the eye. By tracking the progressive changes in capillary flow and microvascular structure over the course of diabetes from weeks- to-years, we seek to better understand the earliest events leading to vascular disease of the eye. In this study, we examine the impact of high blood sugar levels on a mouse model of human diabetes. Changes in single- cell blood flow will be non-invasively imaged over time to determine the impact of diabetes on the smallest vessels of the eye.

总结/摘要 在美国，糖尿病视网膜病变是工作年龄成人失明的主要原因， 全世界的公共卫生问题。这种眼病的最早表现被认为是 起源于毛细血管功能障碍，导致局部毛细血管的过度灌注和灌注不足。虽然 这些微血管结构的变化已被确定为疾病的标志，最早的 这个微观网络的功能变化仍不清楚。微血管血流是否在 毛细血管结构的变化，或者异常血管模式的形成，作为结果， 改变视网膜毛细血管的流动 传统的视网膜照相机一般缺乏必要的分辨率来研究毛细血管水平的血流 因为眼睛的光学系统模糊了眼睛后部的毛细血管。在这项研究中，我们开发和 部署一种新的视网膜摄像机，将眼睛变成一个高倍显微镜，以研究单细胞血流， 活眼的背后结合自适应光学相机的光学改进， 对于图像模糊，我们结合了另外两项创新，以成像单个血细胞的运动，因为它们 通过最微小的毛细血管只有人类头发厚度的十分之一。首先，血细胞不仅 微观的，但它们也以很快的速度移动。为了对这些血细胞进行无运动模糊的成像，使用 需要高速摄像机。在这项研究项目中，我们将联合收割机的模糊校正光学与 超高速相机，每秒可拍摄超过30，000张快照。这台相机的焦点是 单个毛细血管，并可以成像血细胞，因为他们流动-一个接一个。这一进步使我们能够 测量血细胞速度，并提供通过血细胞数量的准确计数，两个创新 测量毛细血管水平的血流。第二个创新是使用特殊的光散射特性， 血细胞，提供血管壁和周围血细胞边界的高度详细图像 组织.所得到的图像不仅提供血细胞的高分辨率图像，而且还可以提供 前所未有的测量血细胞类型及其在眼睛微血管内的变形。通过跟踪 糖尿病病程中毛细血管流动和微血管结构的进行性变化， 年，我们寻求更好地了解导致眼部血管疾病的最早事件。在本研究中， 我们研究了高血糖水平对人类糖尿病小鼠模型的影响。单方面的变化- 细胞血流将随着时间的推移进行非侵入性成像，以确定糖尿病对最小细胞的影响。 眼睛的血管