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Shared TIRF and Patterned Illumination Fluorescence Microscope

共享 TIRF 和图案照明荧光显微镜

基本信息

批准号：
7792945
负责人：
Aaron F Straight
金额：
$ 46.49万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-06-10 至 2011-06-09
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): This application requests funds to purchase a multi-use fluorescence microscope to be shared by three NIH R01 supported Principal Investigators and one NIH K99 supported investigator in the Department of Biochemistry at Stanford University School of Medicine. The laboratories of Jim Spudich, Suzanne Pfeffer, Aaron Straight and Rajat Rohatgi share a common interest in microscopy based dynamic analysis of cellular systems and mechanistic dissection of those systems through biochemical analysis. Central to the research in our laboratories is the use of fluorescence microscopy to study cellular protein and organelle dynamics. In order to analyze complex cell biological processes, our laboratories combine optical, chemical, and genetic perturbation of cells with high-resolution immunofluorescence imaging and live cell timelapse imaging. To understand how individual cellular components participate in cellular functions we apply microscopy-based analysis to reconstituted biochemical systems and cell extract based reactions that recapitulate cellular processes. Specifically, we use methods ranging from single molecule analysis of motor protein movement on cytoskeletal polymers by optical trapping and TIRF microscopy to epifluorescence imaging of chromosome segregation and vesicle trafficking in cell extracts. We are requesting funds to purchase a shared microscope capable of live cell time-lapse fluorescence and brightfield microscopy, TIRF microscopy, patterned fluorescence photobleaching and photoactivation and iterative deconvolution. These combined modalities will significantly enhance the NIH supported research in our laboratories by providing a shared resource to address our microscopy needs.

描述（由申请人提供）：本申请要求资金购买一台多用途荧光显微镜，由斯坦福大学医学院生物化学系的三名 NIH R01 支持的首席研究员和一名 NIH K99 支持的研究人员共享。 Jim Spudich、Suzanne Pfeffer、Aaron Straight 和 Rajat Rohatgi 的实验室对基于显微镜的细胞系统动态分析以及通过生化分析对这些系统进行机械解剖有着共同的兴趣。我们实验室研究的核心是使用荧光显微镜来研究细胞蛋白质和细胞器动力学。为了分析复杂的细胞生物过程，我们的实验室将细胞的光学、化学和遗传扰动与高分辨率免疫荧光成像和活细胞延时成像相结合。为了了解单个细胞成分如何参与细胞功能，我们将基于显微镜的分析应用于重建的生化系统和基于细胞提取物的反应，以概括细胞过程。具体来说，我们使用的方法包括通过光学捕获和 TIRF 显微镜对细胞骨架聚合物上的运动蛋白运动进行单分子分析，以及细胞提取物中染色体分离和囊泡运输的落射荧光成像。我们正在申请资金购买一台能够进行活细胞延时荧光和明场显微镜、TIRF 显微镜、图案荧光光漂白和光激活以及迭代解卷积的共享显微镜。这些组合模式将通过提供共享资源来满足我们的显微镜需求，从而显着增强 NIH 支持的我们实验室的研究。