A DECONVOLUTION MICROSCOPE FOR CELL BIOLOGICAL RESEARCH

用于细胞生物学研究的解卷积显微镜

• 批准号: 6291344
• 负责人: Mark David Rose
• 金额: $ 22.69万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-15 至 2003-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6291344
• 关键词: biomedical equipment purchase; gene expression; microscopy; protein localization

We request funding for the purchase of an integrated digital deconvolution microscope workstation to examine the expression and localization of proteins, chromosomes, organelles, viruses and cytoskeletal elements in fixed and living cells. With the new digital deconvolution microscope, researchers at Princeton University will be able to optically section biological specimens with unprecedented resolution (better than 0.2 micron) and clarity. The deconvolution microscope can achieve this outstanding level of resolution by using computational methods to digitally remove "out-of focus" light from other focal planes. The optical slices can them be combined to produce highly accurate three-dimensional models of sub-cellular structures. Coupled to fast and extremely sensitive digital cameras (able to detect single photons), the deconvolution microscope will allow researchers to follow rapid dynamic processes within living cells using fast time-lapse imaging. This proposal details four major, and one minor, projects where the digital deconvolution will have a major impact. These projects include: (1) elucidation of the long range chromosome interactions that occur during site-specific mitotic recombination such as occurs during mating type interconversion in yeast, (2) the visualization of Herpes virus invasion and spreading in neuronal cells, (3) analyzing the roles of microtubule orientation and dynamics on nuclear orientation and migration using yeast as a model system, (4) visualization of telomeres and the nuclear envelope to examine the role of nuclear peripheral localization on the transcriptional repression that occurs at telomeres, and (5) analysis of the secretory pathway in live yeast cells to test different models of Golgi biogenesis. In each case, because of the extremely small size of the cells and/or of their internal structures, these projects require the unique optical sectioning power and resolution of the digital deconvolution microscope. By these means, the digital deconvolution microscope will provide outstanding research and educational opportunities on one of the most powerful optical microscopes available for use in the life sciences.

我们要求购买综合数字反卷积显微镜工作站，以检查固定和活细胞中蛋白质，染色体，细胞器，病毒和细胞骨架元素的表达和定位。借助新的数字反卷积显微镜，普林斯顿大学的研究人员将能够以前所未有的分辨率（大于0.2微米）和清晰度来将生物标本视为分截面。反卷积显微镜可以通过使用计算方法来数字地删除其他焦点平面的“焦点外”光，从而实现了这种出色的分辨率。可以将光学切片组合在一起以产生高准确的三维次细胞结构模型。反卷积显微镜将结合到快速且极其敏感的数字摄像机（能够检测单个光子），将使研究人员能够使用快速的延时成像来遵循活细胞内的快速动态过程。该提案详细介绍了数字反卷积将产生重大影响的四个主要和一个未成年人项目。这些项目包括：（1）阐明在酵母中交配类型相互转换期间发生的远距离染色体相互作用，例如发生在酵母中的交配类型相互转换期间，（2）（2）使用微管型和动态化的核定型在神经元细胞中的可视化和神经元细胞中的传播，（3）分析i型型型型型型型型和动力学的迁移（3）端粒和核包膜检查核周围定位在端粒中发生的转录抑制作用中的作用，以及（5）分泌途径在活酵母细胞中分析以测试高尔基生物发生模型的分泌途径。在每种情况下，由于细胞的大小和/或它们的内部结构的尺寸极小，这些项目需要数字反卷积显微镜的独特光学切片功率和分辨率。通过这些方式，数字反卷积显微镜将为可用于生命科学的最强大的光学显微镜之一提供出色的研究和教育机会。