Quantitative Fluorescent Speckle Microscopy

定量荧光散斑显微镜

基本信息

  • 批准号:
    6991224
  • 负责人:
  • 金额:
    $ 40.63万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2003
  • 资助国家:
    美国
  • 起止时间:
    2003-01-01 至 2007-09-30
  • 项目状态:
    已结题

项目摘要

DESCRIPTION (provided by applicant): We have developed a powerful new Fluorescent Speckle Microscopy (FSM) method that utilizes epi-fluorescence microscopy to analyze the movement, assembly, and disassembly of macromolecular structures in vivo and in vitro. Initially, we devised this method for studying the dynamics of the cytoskeletal polymers, microtubules and actin filaments in cell locomotion and division. In FSM, structures are assembled from a low fraction of fluorescently labeled subunits together with unlabeled subunits and imaged with high-resolution optics and a sensitive, low-noise digital camera. FSM is achieved in living cells by microinjection or expression of a low amount of fluorophore-conjugated subunits. Stochastic variations in the number of fluorescent subunits per resolution-limited image region results in a "speckled" appearance of the assembled structure. In time-lapse FSM, movement and changes in speckle intensity act as reporters for structure translocation, assembly and disassembly. FSM is fast becoming the method of choice for studies of cytoskeleton dynamics. However, FSM requires further development to reach its full potential for biomedical research. Focus drifts during imaging are a major source of FSM artifact, and the versatility of FSM for various fluorescence microscope modes has to be demonstrated. In addition, the analysis of time-lapse FSM images are currently done by hand, which is slow, incomplete, and prone to error. This must be replaced by automated, quantitative FSM image analysis. Finally, FSM image generation and analysis techniques have to be broadened to general biomacromolecular assemblies. To achieve these goals, we have the following Specific Aims: 1. Hardware Development: To develop focus-stabilized FSM and multi-spectral Total Internal Reflection FSM (TIR-FSM) for studying protein dynamics in vitro and on ventral cell surfaces in vivo. 2. Software Development: To develop software for automatic quantitative analysis of protein dynamics in time-lapse FSM images. 3. Applications: To use multi-spectral FSM, TIR-FSM, and analysis software for studying other biologically important systems besides cytoskeletal polymers. These advances will allow FSM to become a powerful tool for studying molecular dynamics in cell biology, neurobiology, biotechnology, and material science.
描述(由申请人提供):

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

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Clare Michal Waterman其他文献

Clare Michal Waterman的其他文献

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{{ truncateString('Clare Michal Waterman', 18)}}的其他基金

NIH Director's Pioneer Award
NIH 院长先锋奖
  • 批准号:
    7271235
  • 财政年份:
    2005
  • 资助金额:
    $ 40.63万
  • 项目类别:
NIH Director's Pioneer Award (RMI)
NIH 院长先锋奖 (RMI)
  • 批准号:
    7079010
  • 财政年份:
    2005
  • 资助金额:
    $ 40.63万
  • 项目类别:
ASCB Summer Meeting: Coordinating the Events of Directed
ASCB 夏季会议:协调指导活动
  • 批准号:
    7000744
  • 财政年份:
    2005
  • 资助金额:
    $ 40.63万
  • 项目类别:
NIH Director's Pioneer Award
NIH 院长先锋奖
  • 批准号:
    7128506
  • 财政年份:
    2005
  • 资助金额:
    $ 40.63万
  • 项目类别:
Quantitative Fluorescent Speckle Microscopy
定量荧光散斑显微镜
  • 批准号:
    6839430
  • 财政年份:
    2003
  • 资助金额:
    $ 40.63万
  • 项目类别:
Quantitative Fluorescent Speckle Microscopy
定量荧光散斑显微镜
  • 批准号:
    6569910
  • 财政年份:
    2003
  • 资助金额:
    $ 40.63万
  • 项目类别:
Quantitative Fluorescent Speckle Microscopy
定量荧光散斑显微镜
  • 批准号:
    6693772
  • 财政年份:
    2003
  • 资助金额:
    $ 40.63万
  • 项目类别:
MICROTUBULE/ACTIN INTERACTIONS IN CELL MOTILITY
细胞运动中的微管/肌动蛋白相互作用
  • 批准号:
    6181997
  • 财政年份:
    2000
  • 资助金额:
    $ 40.63万
  • 项目类别:
Microtubule/Actin Interactions in Cell Motility
细胞运动中的微管/肌动蛋白相互作用
  • 批准号:
    7118202
  • 财政年份:
    2000
  • 资助金额:
    $ 40.63万
  • 项目类别:
MICROTUBULE/ACTIN INTERACTIONS IN CELL MOTILITY
细胞运动中的微管/肌动蛋白相互作用
  • 批准号:
    6525938
  • 财政年份:
    2000
  • 资助金额:
    $ 40.63万
  • 项目类别:

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定量荧光散斑显微镜
  • 批准号:
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  • 财政年份:
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    $ 40.63万
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定量荧光散斑显微镜
  • 批准号:
    8135496
  • 财政年份:
    2003
  • 资助金额:
    $ 40.63万
  • 项目类别:
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  • 批准号:
    9752630
  • 财政年份:
    2003
  • 资助金额:
    $ 40.63万
  • 项目类别:
Quantitative Fluorescent Speckle Microscopy
定量荧光散斑显微镜
  • 批准号:
    6569910
  • 财政年份:
    2003
  • 资助金额:
    $ 40.63万
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定量荧光散斑显微镜
  • 批准号:
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  • 财政年份:
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    $ 40.63万
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定量荧光散斑显微镜
  • 批准号:
    8015799
  • 财政年份:
    2003
  • 资助金额:
    $ 40.63万
  • 项目类别:
Quantitative Fluorescent Speckle Microscopy
定量荧光散斑显微镜
  • 批准号:
    8255920
  • 财政年份:
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    $ 40.63万
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Quantitative Fluorescent Speckle Microscopy
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