Spinning disk confocal / FRAP microscope for quantitative live cell imaging

用于定量活细胞成像的转盘共焦/FRAP 显微镜

基本信息

项目摘要

DESCRIPTION (provided by applicant): We are requesting funds to purchase a state-of-the-art Spinning Disk Confocal Microscope System specifically designed for live cell imaging of mammalian cells and tissues. The instrument will be used by a core group of seven NIH-funded investigators and their laboratories to facilitate fourteen NIH- funded projects. The principle types of experiments for which this microscope system will be used include multidimensional imaging of fluorescently tagged proteins in living cells and tissues, analysis of protein activity by FRET, analysis of protein and organelle dynamics by FRAP or photoactivation, and photoablation or release of caged compounds in living cells or tissues. The system we are requesting includes a Yokogawa CSU-X1 spinning disk scanner on a Nikon Ti-E inverted microscope stand, which has a built-in focus feedback system allowing imaging for many hours at high resolution with no discernable focus drift. The microscope will also be equipped with an environmental chamber, a motorized stage, and CCD cameras for high resolution and high sensitivity imaging. Oil- and water- immersion objectives are requested for the optimal collection of data from live cells or tissues in a 2D or 3D environment at the highest achievable optical resolution. Solid-state laser lines are requested at 440, 488, 514, and 561 nm to permit imaging of all commonly used fluorescent protein variants. In addition, this instrument will be equipped with a pulsed dye laser-based photobleaching / photoablation module extending its usefulness beyond simple observation of live specimens. Currently, no spinning disk microscope system exists at UCSF that has FRAP or photoactivation capabilities, which severely limits the types of experiments that can be performed. It is expected that the major users will utilize the instrument for ~80% time. However, because the instrument will be housed in and managed through the UCSF Parnassus Biological Imaging Development Center, training and use will be available to additional NIH-funded investigators within the UCSF community.
描述(申请人提供):我们正在申请资金购买最先进的旋转圆盘共聚焦显微镜系统,专门为哺乳动物细胞和组织的活细胞成像而设计。该仪器将由七名NIH资助的研究人员及其实验室组成的核心小组使用,以促进NIH资助的14个项目。该显微镜系统将用于的主要实验类型包括:活细胞和组织中荧光标记蛋白质的多维成像,FRET分析蛋白质活性,FRAP或光激活分析蛋白质和细胞器动力学,以及活细胞或组织中笼子化合物的光消融或释放。我们要求的系统包括安装在尼康Ti-E倒置显微镜支架上的横河CSU-X1旋转圆盘扫描仪,它具有内置的焦点反馈系统,可以在高分辨率下进行长达数小时的成像,而不会出现明显的焦移。显微镜还将配备一个环境室、一个电动工作台和用于高分辨率和高灵敏度成像的CCD摄像机。为了以可达到的最高光学分辨率从2D或3D环境中的活细胞或组织中最佳地收集数据,需要油浸物镜和水浸物镜。要求在440、488、514和561 nm处使用固态激光线路,以允许对所有常用的荧光蛋白质变体进行成像。此外,该仪器将配备基于脉冲染料激光的光漂白/光消融模块,将其用途扩展到简单地观察活标本之外。目前,加州大学旧金山分校不存在具有FRAP或光激活功能的旋转磁盘显微镜系统,这严重限制了可以进行的实验类型。预计主要用户将使用该仪器约80%的时间。然而,由于该仪器将被安置在加州大学旧金山分校帕纳苏斯生物成像开发中心并通过其进行管理,加州大学洛杉矶分校社区内更多的NIH资助的研究人员将获得培训和使用。

项目成果

期刊论文数量(5)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Basolateral junction proteins regulate competition for the follicle stem cell niche in the Drosophila ovary.
  • DOI:
    10.1371/journal.pone.0101085
  • 发表时间:
    2014
  • 期刊:
  • 影响因子:
    3.7
  • 作者:
    Kronen MR;Schoenfelder KP;Klein AM;Nystul TG
  • 通讯作者:
    Nystul TG
Enzymatically Generated CRISPR Libraries for Genome Labeling and Screening.
酶促生成的CRISPR库用于基因组标记和筛选。
  • DOI:
    10.1016/j.devcel.2015.06.003
  • 发表时间:
    2015-08-10
  • 期刊:
  • 影响因子:
    11.8
  • 作者:
    Lane AB;Strzelecka M;Ettinger A;Grenfell AW;Wittmann T;Heald R
  • 通讯作者:
    Heald R
Methods for Imaging Intracellular pH of the Follicle Stem Cell Lineage in Live Drosophila Ovarian Tissue.
活果蝇卵巢组织中卵泡干细胞谱系的细胞内 pH 值成像方法。
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Torsten Wittmann其他文献

Torsten Wittmann的其他文献

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{{ truncateString('Torsten Wittmann', 18)}}的其他基金

Probing Microtubule Function in Neuronal Development
探索神经元发育中的微管功能
  • 批准号:
    10116503
  • 财政年份:
    2018
  • 资助金额:
    $ 50万
  • 项目类别:
Probing Microtubule Function in Neuronal Development
探索神经元发育中的微管功能
  • 批准号:
    10362567
  • 财政年份:
    2018
  • 资助金额:
    $ 50万
  • 项目类别:
Probing Microtubule Function in Neuronal Development
探索神经元发育中的微管功能
  • 批准号:
    9886299
  • 财政年份:
    2018
  • 资助金额:
    $ 50万
  • 项目类别:
Light-activated proteolysis as a tool to analyze intracellular protein function
光激活蛋白水解作为分析细胞内蛋白质功能的工具
  • 批准号:
    8539033
  • 财政年份:
    2010
  • 资助金额:
    $ 50万
  • 项目类别:
Light-activated proteolysis as a tool to analyze intracellular protein function
光激活蛋白水解作为分析细胞内蛋白质功能的工具
  • 批准号:
    8325134
  • 财政年份:
    2010
  • 资助金额:
    $ 50万
  • 项目类别:
Light-activated proteolysis as a tool to analyze intracellular protein function
光激活蛋白水解作为分析细胞内蛋白质功能的工具
  • 批准号:
    7993343
  • 财政年份:
    2010
  • 资助金额:
    $ 50万
  • 项目类别:
Light-activated proteolysis as a tool to analyze intracellular protein function
光激活蛋白水解作为分析细胞内蛋白质功能的工具
  • 批准号:
    8132228
  • 财政年份:
    2010
  • 资助金额:
    $ 50万
  • 项目类别:
Microtubule dynamics during cell polarity and migration
细胞极性和迁移过程中的微管动力学
  • 批准号:
    7808914
  • 财政年份:
    2008
  • 资助金额:
    $ 50万
  • 项目类别:
Microtubule dynamics during cell polarity and migration
细胞极性和迁移过程中的微管动力学
  • 批准号:
    8989112
  • 财政年份:
    2008
  • 资助金额:
    $ 50万
  • 项目类别:
Microtubule dynamics during cell polarity and migration
细胞极性和迁移过程中的微管动力学
  • 批准号:
    7614315
  • 财政年份:
    2008
  • 资助金额:
    $ 50万
  • 项目类别:

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