A Spectral/Lifetime Microscope for in vivo Studies

用于体内研究的光谱/寿命显微镜

• 批准号: 7256572
• 负责人: John Graham White
• 金额: $ 0.68万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7256572
• 关键词: Caenorhabditis elegans; bioimaging /biomedical imaging; biological models; biomedical equipment development; cheek pouch technique; clinical research; collagen; computer human interaction; computer program /software; computer system design /evaluation; dimethylbenzanthracene; electromagnetic radiation; embryo /fetus; extracellular matrix; fluorescence microscopy; fluorescent dye /probe; hamsters; ionophores; keratinocyte; mathematical model; neoplasm /cancer diagnosis; neoplastic cell culture for noncancer research; photomultiplier; radiation detector; squamous cell carcinoma; transfection

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，可以深入了解固定标本观察无法获得的动态行为和细胞生理学。这些信息可以向医生揭示组织的病理状态，或者向研究科学家揭示细胞骨架动力学的信息。全基因组测序与荧光蛋白报告基因的使用相结合，开辟了可视化活组织中任何蛋白质的分布和动态的可能性。此外，特异性荧光指示剂的使用允许在体内可视化细胞的生理状态。然而，重叠的光谱，低信号和光散射的问题造成了严重的限制，在实践中可以实现与荧光显微镜。这些问题将通过开发多光子成像系统来解决，该系统最大限度地利用荧光信号中存在的所有信息。一种新型的光子计数探测器将被开发，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提取噪声限制信号中单个荧光成分的物理意义特征。该系统将被应用于细胞生物学研究，使用荧光报告，并确定潜在的诊断光谱/寿命签名的细胞和组织培养模型的癌症。

项目成果

Applications of combined spectral lifetime microscopy for biology.

组合光谱寿命显微镜在生物学中的应用。

• DOI: 10.2144/000112251
• 发表时间: 2006
• 期刊: BioTechniques
• 影响因子: 2.7
• 作者: Yan,Long;Rueden,CurtisT;White,JohnG;Eliceiri,KevinW
• 通讯作者: Eliceiri,KevinW

Multiphoton redox ratio imaging for metabolic monitoring in vivo.

• DOI: 10.1007/978-1-60761-411-1_11
• 发表时间: 2010
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Skala, Melissa;Ramanujam, Nirmala
• 通讯作者: Ramanujam, Nirmala

Nonlinear optical microscopy and computational analysis of intrinsic signatures in breast cancer.

• DOI: 10.1109/iembs.2009.5334523
• 发表时间: 2009
• 期刊: Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Annual International Conference
• 作者: Rueden CT;Conklin MW;Provenzano PP;Keely PJ;Eliceiri KW
• 通讯作者: Eliceiri KW

Collagen density promotes mammary tumor initiation and progression.

• DOI: 10.1186/1741-7015-6-11
• 发表时间: 2008-04-28
• 期刊: BMC medicine
• 影响因子: 9.3
• 作者: Provenzano PP;Inman DR;Eliceiri KW;Knittel JG;Yan L;Rueden CT;White JG;Keely PJ
• 通讯作者: Keely PJ

Fast localized wavefront correction using area-mapped phase-shift interferometry.

使用区域映射相移干涉测量法进行快速局部波前校正。

• DOI: 10.1364/ol.36.002892
• 发表时间: 2011
• 期刊: Optics letters
• 影响因子: 3.6
• 作者: Hall,Gunnsteinn;Spalding,GabrielC;Campagnola,PaulJ;White,JohnG;Eliceiri,KevinW
• 通讯作者: Eliceiri,KevinW