A Neurolucida and Stereo Investigator Imaging System with MicroLucida

采用 MicroLucida 的 Neurolucida 和 Stereo Investigator 成像系统

• 批准号: 8247678
• 负责人: FRANK M LAFERLA
• 金额: $ 31.08万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-15 至 2013-05-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8247678
• 关键词: Alzheimer' s Disease; Astrocytes; Biological; Brain; Cell Count; Cell Transplants; Cicatrix; Cognitive; Dimensions; Disease; Engraftment; Etiology; Funding; Gliosis; Huntington Disease; Image; Imaging Techniques; Impairment; Injury; Institutes; Label; Lesion; Lighting; Measurement; Memory impairment; Methods; Motor; Multiple Sclerosis; Nervous system structure; Neurodegenerative Disorders; Neurons; Research Personnel; Science; Senile Plaques; Spinal cord injury; Structure; Synapses; System; Three-Dimensional Image; Time; Tissues; Traumatic Brain Injury; Ventricular; base; brain irradiation injury; developmental disease; immunocytochemistry; member; nervous system disorder; neurogenesis; neuroinflammation; neuron loss; relating to nervous system; statistics; tissue processing

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).

描述(申请人提供)：阿尔茨海默氏症和亨廷顿病、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等神经疾病的特征是严重的神经元和神经胶质损伤，导致认知和/或运动功能严重受损。虽然这些疾病的病因各不相同，但许多疾病的标志性和决定性的病理特征是神经元和突触的丧失，神经元连通性的丧失，组织体积的丧失和脑室空间的扩张，神经发生的失调，以及猖獗的神经炎症、胶质增生和瘢痕形成。该提案中列出的研究人员都是加州大学欧文分校记忆损伤和神经疾病研究所(UCI Mind)的成员，他们正在寻求资金，以获得一种多通道荧光结构照明立体成像系统，以无偏见的方法量化组织变化。神经病理学研究严重受限于试图通过二维观察来研究三维相互作用。然而，体视学是一门通过提供生物组织平面切片的三维解释来克服这些限制的科学，因为它是基于几何学和统计学原理的融合。因此，神经系统的体视学研究提供了一种对细胞数量、神经突起和组织体积进行更精确的定量测量的方法。此外，对于神经退行性疾病的研究，例如，有可能确定大脑损伤(如淀粉样斑块)的积累和细胞数量之间的定量关系。标准的Brightfield体视学使人们可以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门的成像技术。Apotome结构化照明系统允许获取共焦样3D图像以用于多个标签的实时和离线分析。这样的系统将允许研究人员同时量化两到三个参数(例如，表达GFP的移植细胞总数，以及第二个或第三个命运标记，以获得同一组织切片中神经、星形胶质细胞和少突胶质细胞命运的总植入数量和百分比)。

项目成果

Inhibition of hematopoietic cell kinase dysregulates microglial function and accelerates early stage Alzheimer's disease-like neuropathology.

• DOI: 10.1002/glia.23522
• 发表时间: 2018-12
• 期刊: Glia
• 影响因子: 6.2
• 作者: Lim SL;Tran DN;Zumkehr J;Chen C;Ghiaar S;Kieu Z;Villanueva E;Gallup V;Rodriguez-Ortiz CJ;Kitazawa M
• 通讯作者: Kitazawa M