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

• 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
• 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

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.

项目概要 尽管 2 型糖尿病对周围器官的小血管影响相当好 据了解，糖尿病对脑微血管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是技术挑战 与体内小血管网络拓扑的完整三维映射相关。 技术挑战在于在毛细管尺度上捕获血管形态的能力，同时 也跨越整个皮质厚度并延伸到皮质下区域。更远 使技术挑战变得更加复杂的是，船舶网络的基线差异很大 动物的结构和对病理扰动的反应，因此血管网络必须 纵向跟踪以捕捉这些动态。因此，本提案的目标是 通过开发新的光学显微镜技术来克服这些技术限制 体内整个皮质微血管网络结构的完整3D映射，并使用 这些技术可以量化与类型相关的脑微血管系统的慢性变化 2 小鼠糖尿病。我们将介绍二光子和三光子荧光的新进展 显微镜将显着提高我们体内微血管网络成像的能力 在小鼠皮层的大空间范围内深度可达 1.5 毫米。我们将开发一个新的空间 偏移双色多光子激发方法利用长波长的最新进展， 同步超快激光器。皮质和皮质下脉管系统的大规模图像将 允许血管网络的完整 3D 矢量化，从而能够对血管网络进行详细分析 随时间变化的微血管形态。我们将在体内使用这种先进的组合 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血管结构变化。