A Spectral/Lifetime Microscope for in vivo Studies

用于体内研究的光谱/寿命显微镜

基本信息

批准号：
7121008
负责人：
John Graham White
金额：
$ 3.18万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-08-01 至 2006-05-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Observations of living cells or tissues through fluorescence microscopy provide insights into dynamic behavior and cellular physiology that cannot be obtained from observations of fixed specimens. Such information may reveal the pathological state of a tissue to the physician or information on cytoskeletal dynamics to the research scientist. Whole genome sequencing combined with the use of fluorescent protein reporters has opened up the possibility of visualizing the distribution and dynamics of any protein in living tissue. Furthermore, the use of specific fluorescent indicators has allowed the physiological state of a cell to be visualized in vivo. However, problems of overlapping spectra, low signal and light scatter pose serious limitations on what can be achieved in practice with fluorescence microscopy. These problems will be addressed by the development of a multi-photon imaging system that makes maximum use of all the information present in the fluorescence signal. A novel photon-counting detector will be developed that simultaneously measures the wavelength and lifetime of individual fluorescent photons. Algorithms will be developed to analyze and extract physically meaningful characteristics of individual fluorescent components from noise-limited signals. The system will be applied to cell biological studies using fluorescent reporters and to the identification of potentially diagnostic spectral/lifetime signatures of cells and tissue culture models of cancer.

描述（由申请人提供）：通过荧光显微镜观察活细胞或组织的观察提供了对动态行为和细胞生理学的见解，这些观察是从固定标本的观察结果中无法获得的。这样的信息可能会揭示组织对医师的病理状态，或者向研究科学家提供有关细胞骨架动力学的信息。整个基因组测序与荧光蛋白报道的使用结合使用，已经开辟了可视化活组织中任何蛋白质的分布和动力学的可能性。此外，使用特定的荧光指标使细胞的生理状态在体内可视化。但是，重叠光谱，低信号和光散射的问题对荧光显微镜实践中可以实现的目标产生了严重的限制。这些问题将通过开发多光子成像系统的开发来解决，该系统最大程度地利用了荧光信号中存在的所有信息。将开发出一种新型的光子计数检测器，同时测量单个荧光光子的波长和寿命。将开发算法来分析和从噪声限制信号中分析和提取单个荧光成分的物理有意义的特征。该系统将用于使用荧光记者的细胞生物学研究，并鉴定细胞和癌症的组织培养模型的潜在诊断光谱/寿命特征。