Longitudinal Imaging of Cortical Small Vessel Network Structure with Two-Color Multiphoton Fluorescence Microscopy

双色多光子荧光显微镜对皮质小血管网络结构的纵向成像

基本信息

批准号：
10217270
负责人：
Andrew K Dunn
金额：
$ 54.32万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10217270
关键词：
3-Dimensional Adopted Affect Anatomy Animals Blood Vessels Blood capillaries Body part Brain Brain Diseases Cerebrum Chronic Color Complex Complications of Diabetes Mellitus Computer Analysis Diabetes Mellitus Diabetic Retinopathy Disease Fluorescence Microscopy Functional disorder Goals Heart Diseases Histologic Image Knowledge Lasers Maps Methods Microscope Microscopy Morphology Multiphoton Fluorescence Microscopy Mus Myocardium Neurodegenerative Disorders Neurologic Non-Insulin-Dependent Diabetes Mellitus Optics Organ Pathologic Penetration Peripheral Photons Resolution Retina Risk Rodent Model Role Signal Transduction Speed Structure System Techniques Thick Time Tissues Vascular Diseases Vascular System Work base cerebral microvasculature contrast imaging design fluorescence imaging imaging modality improved in vivo in vivo imaging insight instrumentation multiphoton imaging multiphoton microscopy neurovascular new technology novel optical imaging response serial imaging tool two-photon vector

项目摘要

Project Summary Although the small vessel consequences of type 2 diabetes in peripheral organs are fairly well understood, the effects of diabetes on cerebral microvasculature are largely unknown. A significant impediment to understanding these changes has been the technical challenges associated with complete three-dimensional mapping of small vessel network topology in vivo. The technical challenge lies in the ability to capture vessel morphology on the capillary scale while also spanning the entire cortical thickness and extending into the subcortical regions. Further complicating the technical challenge is the fact that vessel networks vary significantly in baseline structure and response to pathological perturbations across animals, thus vessel networks must be followed longitudinally to capture these dynamics. Therefore, the goal of this proposal is to overcome these technical limitations by developing new optical microscopy techniques that permit complete 3D mapping of the entire cortical microvascular network structure in vivo, and to use these techniques to quantify chronic alterations in cerebral microvasculature associated with type 2 diabetes in mice. We will introduce new advances to two- and three-photon fluorescence microscopy that will significantly improve our ability to image microvascular networks in vivo at depths up to 1.5 mm over large spatial extents of mouse cortex. We will develop a new spatially offset two-color multiphoton excitation approach that utilizes recent advances in long wavelength, synchronized ultrafast lasers. The large-scale images of cortical and sub-cortical vasculature will allow full 3D vectorization of vascular networks that will enable detailed analysis of the microvascular morphology over time. We will use this combination of advanced in vivo microscopy, novel ultrafast lasers, and computational analysis of vascular networks to quantify the structural changes in cerebral microvasculature associated with type 2 diabetes.

项目摘要尽管外围器官中2型糖尿病的小血管后果相当不错理解，糖尿病对脑微脉管系统的影响在很大程度上尚不清楚。一个理解这些变化的重大障碍一直是技术挑战与体内小容器网络拓扑的完整三维映射有关。技术挑战在于能够捕获毛细管尺度的船只形态的能力，而还跨越整个皮质厚度并延伸到皮层下区域。更远使技术挑战复杂化的事实是，船只网络在基线中差异很大跨动物的病理扰动的结构和反应，因此血管网络必须纵向遵循以捕获这些动态。因此，该提议的目的是通过开发允许的新光学显微镜技术来克服这些技术限制在体内完成整个皮质微血管网络结构的完整3D映射，并使用这些技术来量化与类型相关的脑微脉管系统中的慢性变化小鼠2糖尿病。我们将向两光子和三光子荧光介绍新的进步显微镜将显着提高我们在体内成像微血管网络的能力在小鼠皮层的大空间范围内，深度高达1.5毫米。我们将在空间上开发新的偏移两色多光激励方法，该方法利用了长波长的最新进展，同步超快激光器。皮质和亚皮质脉管系统的大规模图像将允许血管网络的完整3D矢量化，以详细分析随着时间的流逝，微血管形态。我们将使用这种高级体内的组合显微镜，新型超快激光器和血管网络的计算分析以量化与2型糖尿病相关的脑微脉管系统的结构变化。