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.

我们请求资金购买集成数字解卷积显微镜工作站，以检查固定细胞和活细胞中蛋白质、染色体、细胞器、病毒和细胞骨架元件的表达和定位。借助新型数字反卷积显微镜，普林斯顿大学的研究人员将能够以前所未有的分辨率（优于 0.2 微米）和清晰度对生物样本进行光学切片。反卷积显微镜可以通过使用计算方法以数字方式消除其他焦平面的“失焦”光来实现这种出色的分辨率水平。光学切片可以组合起来产生亚细胞结构的高精度三维模型。与快速且极其灵敏的数码相机（能够检测单光子）相结合，反卷积显微镜将使研究人员能够使用快速延时成像来跟踪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项目和一个次要项目。这些项目包括：(1) 阐明位点特异性有丝分裂重组过程中发生的长程染色体相互作用，例如酵母交配型互变过程中发生的长程染色体相互作用，(2) 疱疹病毒入侵和在神经元细胞中传播的可视化，(3)使用酵母作为模型系统分析微管取向和动力学对核取向和迁移的作用，（4）端粒和核膜的可视化以检查核外周定位对发生在转录抑制中的作用端粒，以及（5）分析活酵母细胞的分泌途径，以测试不同的高尔基体生物发生模型。在每种情况下，由于细胞和/或其内部结构的尺寸极小，这些项目需要数字反卷积显微镜独特的光学切片能力和分辨率。通过这些方式，数字反卷积显微镜将为生命科学领域最强大的光学显微镜之一提供出色的研究和教育机会。