Acquisition of a TIRF/Widefield fluorescence microscope for cell biology and neur

购买用于细胞生物学和神经学的 TIRF/宽场荧光显微镜

• 批准号: 8447315
• 负责人: BRYON D GROVE
• 金额: $ 35.81万
• 依托单位: UNIVERSITY OF NORTH DAKOTA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-15 至 2014-07-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8447315
• 关键词: Acute; Alzheimer' s Disease; Animals; Applications Grants; Area; Basic Science; Behavior; Calcium; Calcium Channel; Cell membrane; Cellular biology; Centers of Research Excellence; Cholesterol; Core Facility; Data Storage and Retrieval; Development; Disease; Electrons; Electrophysiology (science); Epilepsy; Equipment; Event; Faculty; Fluorescence; Fluorescence Microscopy; Functional Imaging; Functional disorder; Funding; Goals; Health Sciences; Homeostasis; Image; Imaging technology; Inclusion Body Myositis; Infectious Agent; International; Investigation; Laboratories; Lead; Location; Manitoba; Mass Spectrum Analysis; Mediating; Microscope; Minnesota; Modeling; Molecular; Nerve Degeneration; Neurosciences Research; North Dakota; Online Systems; Parkinson Disease; Pathogenesis; Phosphorylation; Positron-Emission Tomography; Process; Radioactivity; Research; Research Infrastructure; Research Personnel; Role; Salivary Glands; Sampling; Schedule; Support Contracts; Surface; System; Technology; Total Internal Reflection Fluorescent; Transgenic Organisms; United States National Institutes of Health; Universities; Work; amyloidogenesis; dopamine transporter; fluorescence imaging; fluorescence microscope; instrumentation; knockout gene; medical schools; meetings; member; molecular dynamics; neuron loss; palmitoylation; time use; trafficking

DESCRIPTION (provided by applicant): The purpose of this Shared Instrumentation Grant application is to obtain a TIRF/widefield fluorescence microscope, the first in North Dakota, to enhance existing NIH-funded research areas and to develop new avenues of research. The long-term goals of the projects requiring this instrumentation are to understand the mechanisms underlying the pathogenesis of several disease states under investigation at the University of North Dakota including Alzheimers¿ disease, Parkinson¿s disease, inclusion body myositis (IBM), epilepsy and many more. The overall aims of these projects are to 1) investigate the role of endosomal/lysosomal function in amyloidogenesis associated with Alzheimer's disease, 2) investigate the role of TRPC calcium channels and dysregulation of calcium homeostasis in dopaminergic neuronal cell death associated with Parkinson¿s disease and salivary gland dysfunction, 3) investigate the role of palmitoylation and phosphorylation in regulating dopamine transporter trafficking and function, and 4) investigate mechanisms underlaying cholesterol mediated neurodegeneration associated with Alzheimer's disease. Both TIRF microscopy, an imaging technology that allows very precise realtime fluorescent imaging of dynamic molecular events at the cell membrane surface, and state of the art widefield fluorescence microscopy are necessary for investigators of the proposed projects to extend their studies. All of the projects are investigating dynamic molecular events within different cellular compartments. TIRF microscopy is essential to meeting the goals of several of these projects because of their focus on molecular processes at the cell membrane or the need for single molecular fluorescence imaging. In addition, all projects require fluorescence microscopy instrumentation configured for quantitative wide field fluorescence imaging of dynamic events over time using multiple channels and multiple sampling locations to provide statistical rigor. These studies cannot be accomplished with existing fluorescence and confocal imaging equipment. The closest TIRF/wide field fluorescent microscope systems to the University of North Dakota are at the University of Manitoba (160 miles) and the University of Minnesota (350 miles). These distances (and an international border), our documented needs, and the inability of others to accommodate our needs underscore the acute need for this equipment. To achieve our research goals, we propose to purchase and operate an Olympus IX81-ZDC2 cellTIRF microscope system. This microscope will be integrated into our already functional imaging core facility in association with other imaging equipment and will take advantage of existing web-based scheduling, technical support, web-based data storage and sharing, and system contract support. Acquisition of this instrumentation will build institutionally on our ver successful COBRE and INBRE models. The research enterprise at the University of North Dakota has grown substantially to almost $130 million in 2011 with recent infrastructure improvements including vast expansions to the School of Medicine and Health Sciences, a neuroscience research building, and new laboratories for micro- PET, confocal, electron and stereological imaging, mass spectrometry, electrophysiology, as well as animal work involving transgenic and gene knockout technologies, infectious agents and radioactivity, and behavior. This TIRF/wide field fluorescence microscope system will advance the basic science research of our faculty members in a wide range of fields and will lead to development of new research areas.

描述（由申请人提供）：本共享仪器补助金申请的目的是获得一个TIRF/宽视场荧光显微镜，这是北达科他州的第一个，以加强现有的NIH资助的研究领域，并开发新的研究途径。需要这种仪器的项目的长期目标是了解北达科他州大学正在调查的几种疾病状态的发病机制，包括阿尔茨海默病，帕金森病，包涵体肌炎（IBM），癫痫等。这些项目的总体目标是：1）研究内体/溶酶体功能在阿尔茨海默病相关淀粉样蛋白生成中的作用，2）研究TRPC钙通道和钙稳态失调在帕金森病相关多巴胺能神经元细胞死亡中的作用。s疾病和唾液腺功能障碍，3）研究棕榈酰化和磷酸化在调节多巴胺转运蛋白运输和功能中的作用，和4）研究与阿尔茨海默病相关的胆固醇介导的神经变性的机制。TIRF显微镜（一种成像技术，可以对细胞膜表面的动态分子事件进行非常精确的实时荧光成像）和最先进的宽视野荧光显微镜对于拟议项目的研究人员扩展他们的研究是必要的。所有这些项目都在研究不同细胞隔室内的动态分子事件。TIRF显微镜是必不可少的，以满足这些项目中的几个目标，因为他们的重点是在细胞膜上的分子过程或需要单分子荧光成像。此外，所有项目都需要荧光显微镜仪器，配置用于使用多通道和多个采样位置随时间推移对动态事件进行定量宽场荧光成像，以提供统计严谨性。这些研究不能用现有的荧光和共焦成像设备完成。距离北达科他州大学最近的TIRF/宽视场荧光显微镜系统位于马尼托巴大学（160英里）和明尼苏达大学（350英里）。 这些距离（和国际边界），我们的记录需求，以及其他人无法满足我们的需求，突出了对这种设备的迫切需求。为了实现我们的研究目标，我们建议购买和操作奥林巴斯IX 81-ZDC 2细胞TIRF显微镜系统。 该显微镜将与其他成像设备一起集成到我们已经运行的成像核心设施中，并将利用现有的基于网络的调度，技术支持，基于网络的数据存储和共享以及系统合同支持。这种仪器的收购将建立在我们非常成功的COBRE和INBRE模型的基础上。北达科他州大学的研究企业在2011年大幅增长到近1.3亿美元，最近的基础设施改善包括对医学和健康科学学院的大规模扩建，神经科学研究大楼，以及微PET，共聚焦，电子和体视成像，质谱，电生理学，以及涉及转基因和基因敲除技术、传染性病原体和放射性以及行为的动物工作。该TIRF/宽视场荧光显微镜系统将推进我们教职员工在广泛领域的基础科学研究，并将导致新的研究领域的发展。