MRI: Acquisition of a Total Internal Reflection Fluorescence (TIRF) Microscope for Research and Education at Marquette University and the Greater Milwaukee Region

MRI：购买全内反射荧光 (TIRF) 显微镜，用于马凯特大学和大密尔沃基地区的研究和教育

• 批准号: 1827883
• 负责人: Lisa Petrella
• 金额: $ 25.14万
• 依托单位: Marquette University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-10-01 至 2021-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1827883&HistoricalAwards=false
• 关键词: MRI; Acquisition; Total; Internal; Reflection

An award is made to support the acquisition of an automated Total Internal Reflection Fluorescence (TIRF) Microscope system to support the research, educational and outreach efforts at Marquette University (MU), Medical College of Wisconsin (MCW) and other research groups in the greater Milwaukee region. TIRF microscopy is an essential tool used to study, with high resolution, the mechanism and dynamics of macromolecular protein complex assembly and movement at cellular surfaces. TIRF has also been extensively adapted to study enzymes at single molecule resolution using various fluorescence-based approaches. The availability of a TIRF microscope at MU and neighboring institutions in the greater Milwaukee areas is required to provide hands-on training to undergraduate, graduate and postdoctoral researchers. The PI and co-PIs will provide training to all interested researchers, including students ranging in expertise from graduate to young undergraduate students. The PI and co-PIs have a strong record of training undergraduate and graduate students and the TIRF microscope will become a key part of the educational efforts at Marquette University and surrounding institutions in the greater Milwaukee region.The projects underway in the laboratories of the PI, Co-PIs and other users fall under two broad categories of TIRF microscopy applications. The first group will use single molecule FRET to visualize the motions of fluorescently labelled enzymes and this information will be deconvoluted to understand their mechanism of action. The second group will utilize the TIRF microscope to study in vivo cellular processes, where the dynamics of fluorescently labeled proteins are followed within the cell. The TIRF microscope will be invaluable to: (1) performing mechanistic studies of ATP-dependent motor proteins; (2) clarifying the role of carrier domain movements in biotin-dependent metabolic enzymes; (3) identifying the assembly mechanism of prion formation on actin networks; (4) exploring the role of temperature in gamete development; (5) understand the molecular and cellular mechanisms of eukaryotic flagella; (6) studying Ras GTPase interactions on cellular membranes (7) describing the mechanism of G protein and &#946-arrestin chemokine receptor signaling; and (8) investigating the role of phosphorylation in &#946-arrestin and E3 ubiquitin ligase recruitment. Results from these studies will be published in peer-reviewed scientific journals, presented at scientific meetings, and used in both training and outreach activities.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

该奖项旨在支持自动全内反射荧光（TIRF）显微镜系统的收购，以支持马奎特大学（MU），威斯康星州医学院（MCW）和大密尔沃基地区的其他研究小组的研究，教育和推广工作。TIRF显微镜是一种用于研究高分辨率大分子蛋白质复合物在细胞表面组装和运动的机制和动力学的重要工具。TIRF也已被广泛适用于使用各种基于荧光的方法以单分子分辨率研究酶。在MU和邻近的机构在更大的密尔沃基地区的TIRF显微镜的可用性是必需的，以提供动手培训的本科生，研究生和博士后研究人员。PI和co-PI将为所有感兴趣的研究人员提供培训，包括从研究生到年轻本科生的专业学生。PI和co-PI在培养本科生和研究生方面有着良好的记录，TIRF显微镜将成为马奎特大学和大密尔沃基地区周边机构教育工作的关键部分。PI、Co-PI和其他用户实验室正在进行的项目属于TIRF显微镜应用的两大类。第一组将使用单分子FRET来可视化荧光标记酶的运动，并将这些信息解卷积以了解它们的作用机制。第二组将利用TIRF显微镜研究体内细胞过程，其中荧光标记蛋白质的动力学在细胞内进行。TIRF显微镜对于：（1）进行ATP依赖性马达蛋白的机制研究将是无价的;（2）阐明载体结构域运动在生物素依赖性代谢酶中的作用;（3）确定肌动蛋白网络上朊病毒形成的组装机制;（4）探索温度在配子发育中的作用;（5）了解真核生物鞭毛的分子和细胞机制;（6）研究细胞膜上的Ras GT3相互作用;（7）描述G蛋白和&#946-抑制蛋白趋化因子受体信号传导的机制;和（8）研究磷酸化在&#946-抑制蛋白和E3泛素连接酶募集中的作用。 这些研究的结果将发表在同行评审的科学期刊上，在科学会议上发表，并用于培训和推广活动。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。