A Neurolucida and Stereo Investigator Imaging System with MicroLucida

描述（由申请人提供）：神经系统疾病，如阿尔茨海默氏病和亨廷顿氏病、脊髓损伤、创伤性脑损伤、多发性硬化症、辐射损伤和脑发育障碍，其特征是严重的神经元和神经胶质损伤，导致认知和/或运动功能的显著损伤。尽管这些疾病的病因各不相同，但许多这些疾病的特征和病理特征是神经元和突触的丧失，神经元连通性的丧失，组织体积的丧失和心室空间的扩张，神经发生失调，猖獗的神经炎症，胶质瘤和瘢痕形成。本提案中列出的研究人员都是加州大学欧文分校记忆障碍和神经系统疾病研究所（UCI MIND）的成员，他们正在寻求资金，以获得多通道荧光结构照明立体成像系统，以无偏方法量化组织变化。神经病理学研究由于试图通过二维观察来研究三维相互作用而受到严重限制。然而，立体学是一门通过提供生物组织平面剖面的三维解释来克服这些局限性的科学，因为它是基于几何学和统计学原理的融合。因此，神经系统的立体学研究为获得细胞数量、神经过程和组织体积的更精确定量测量提供了一种手段。此外，对于神经退行性疾病的研究，可以确定例如淀粉样斑块等脑病变的积累与细胞数量之间的定量关系。标准明场立体成像允许在组织切片中一次准确地定量一个免疫标记。双重和/或三重标记免疫细胞化学通常需要荧光标记和更专业的成像技术。Apotome结构化照明系统可以获得类似共聚焦的3D图像，用于实时和离线分析多个标签。这样的系统将允许研究者同时量化两个或三个参数（例如，表达gfp的移植细胞总数，以及第二或第三个命运标记，以获得同一组织切片中神经、星形胶质和少突胶质细胞命运的总移植数量和百分比）。

DESCRIPTION (provided by applicant): Neurological disorders such as Alzheimer's and Huntington's disease, spinal cord injury, traumatic brain injury, multiple sclerosis, radiation injury, and brain developmental disorders are characterized by profound neuronal and glial injury, leading to significant impairments in cognitive and/or motor functions. Although the etiology of these disorders vary, the signature and defining pathological features for many of these conditions is the loss of neurons and synapses, loss of neuronal connectivity, tissue volume loss and expansion of the ventricular spaces, dysregulated neurogenesis, and rampant neuroinflammation, gliosis, and scarring. The investigators listed on this proposal are all members of the Institute for Memory Impairments and Neurological disorders at UC Irvine (UCI MIND), and are seeking funds to acquire a multichannel fluorescent structured illumination stereological imaging system to quantify tissue changes with unbiased methods. Neuropathological studies are severely limited by attempts to study three-dimensional interactions by observations made in two dimensions. Stereology, however, is a science that overcomes these limitations by furnishing three- dimensional interpretations of planar sections of biological tissues, as it is based on merging of the principles of geometry and statistics. Consequently, stereological studies of the nervous system provide a means to obtain more precise quantitative measurements of cell number and neural processes and tissue volume. In addition, for studies on neurodegenerative disorders, it is possible to determine the quantitative relationship between the accumulation of brain lesions such as amyloid plaques and cell number, for example. Standard brightfield stereology allows one to accurately quantify one immunolabel at a time in tisue sections. Double and/or triple labeling immunocytochemistry typically require fluorescent labeling and more specialized imaging techniques. An Apotome structured illumination system allows confocal-like 3D images to be acquired for both real-time and off-line analysis of multiple labels. Such a system would allow investigators to quantify two or three parameters simultaneously (e.g. total number of GFP-expressing transplanted cells, and a second or third fate marker to obtain total engraftment numbers and percentage of neural, astroglia and oligodendroglial fate in the same tissue sections).