Imaging Neuronal and Capillary Dysfunction Deep in the Rodent Brain in vivo Using 1700 NM Optical Coherence Microscopy and Tracer-Based Kinetics

摘要： 皮质下病理学是衰老、阿尔茨海默病和血管性痴呆的常见特征， 很难在体内用微米分辨率进行研究。光学方法，如双光子显微镜 在微米尺度上对表层皮层进行成像，但这些传统显微镜方法的分辨率 在超过600微米成像深度时迅速降解。标准全脑磁共振成像 (MRI)这些方法还不能提供蜂窝级的分辨率，并且实现起来通常是昂贵的。因此 目前迫切需要直接评估深部皮质和皮质下灌注和细胞损伤的方法， 微观水平，从而弥合现有的表面光学显微镜和宏观之间的差距 显像该提案将开发和应用新颖的光学成像技术和伴随方法 直接研究皮质下（海马和白色物质）细胞和血管的遗传变化， 小鼠疾病模型，而不需要荧光蛋白的转基因表达。我们建议 开发和验证方法，以量化单个毛细管水平的渡越时间分布;联合收割机结合这些 测量神经元细胞活力、髓鞘形成、斑块分布、萎缩的方法;最后， 纵向成像阿尔茨海默氏症小鼠模型中深层皮质和海马损伤的时间过程 这些技术将在临床前实验中产生广泛的影响。 研究治疗和生物标志物的发现，并将推进白色物质损伤的研究， 皮质下痴呆这里提出的初步开发、验证和演示将促进 广泛采用这些新技术，以非侵入性方式研究皮质下病理生理学， 老鼠的大脑

Abstract: Subcortical pathology is a common feature in aging, Alzheimer's disease and vascular dementia but has been extremely difficult to study with micron resolution in vivo. Optical methods such as two-photon microscopy image the superficial cortex at the micron-scale, but the resolution of these conventional microscopic methods degrades rapidly beyond 600 microns imaging depth. Standard whole-brain magnetic resonance imaging (MRI) methods do not yet provide cellular-level resolution and are often expensive to implement. Thus, there is a pressing need for methods to directly assess deep cortical and subcortical perfusion and cellular injury at the microscopic level, thus bridging the gap between existing superficial optical microscopy and macroscopic imaging. This proposal will develop and apply novel optical imaging technologies and accompanying methods to directly investigate subcortical (hippocampal and white matter) cellular and vascular changes in genetic mouse models of disease, without the need for transgenic expression of fluorescent proteins. We propose to develop and validate methods to quantify transit time distribution at the single capillary level; combine these with methods to measure neuronal cell viability, myelination, plaque distribution, atrophy; and finally, to longitudinally image the time course of deep cortical and hippocampal injury in a mouse model of Alzheimer's disease up to a depth of 2 mm. These techniques will have a widespread impact in preclinical experimental research in therapeutics and biomarker discovery, and will advance the study of white matter injury and subcortical dementia. The initial development, validation, and demonstration proposed here will catalyze the widespread adoption of these novel techniques to study subcortical pathophysiology non-invasively in the mouse brain.