Imaging neuronal and capillary dysfunction deep in the rodent brain in vivo using 1700 nm Optical Coherence Microscopy and tracer-based kinetics

 描述（由申请人提供）：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，但在体内用微米分辨率研究是极其困难的。诸如双光子显微镜的光学方法在微米级对表层皮层成像，但是这些常规显微镜方法的分辨率在超过600微米成像深度时迅速降低。标准的全脑磁共振成像（MRI）方法还不能提供细胞级的分辨率，而且实施起来往往很昂贵。因此，迫切需要在微观水平上直接评估深层皮质和皮质下灌注和细胞损伤的方法，从而弥合现有表面光学显微镜和宏观成像之间的差距。该提案将开发和应用新的光学成像技术和相关方法，直接研究遗传性疾病小鼠模型中的皮质下（海马和白色物质）细胞和血管变化，而不需要荧光蛋白的转基因表达。我们建议开发和验证在单个毛细血管水平上量化传输时间分布的方法;联合收割机将这些方法与测量神经元细胞活力、髓鞘形成、斑块分布、萎缩的方法相结合;最后，纵向成像深部皮质的时间过程。 以及阿尔茨海默病小鼠模型中深度达2 mm的海马损伤。这些技术将对治疗学和生物标志物发现的临床前实验研究产生广泛影响，并将推进白色物质损伤和皮质下痴呆的研究。初步的发展，验证和示范提出这里将促进这些新技术的广泛采用，研究皮层下的病理生理非侵入性小鼠大脑。

 DESCRIPTION (provided by applicant): 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.