Zeiss LSM510 META confocal-fluorescence spectroscopy

Zeiss LSM510 META 共焦荧光光谱仪

• 批准号: 6580051
• 负责人: HAROLD P ERICKSON
• 金额: $ 38万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2004-10-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6580051
• 关键词: biochemistry; biomedical equipment purchase; cell biology; confocal scanning microscopy; fluorescence spectrometry

DESCRIPTION (provided by applicant): This application is to purchase a modern laser scanning confocal microscope to serve primarily the needs of the Cell Biology Department, Duke University Medical Center. The only departmental confocal at present is a 10 year old Bio-Rad MRC 600, which has limited capabilities in several areas. The microscope of our choice is the new Zeiss LSM 510 META. The META emission detector provides a unique and powerful detection system that far surpasses conventional dichroic filters. The META detector is based on a diffraction grating that spreads the emission into a full spectrum, and a detector that records the full spectrum from each scanned point. This provides a unique capability to separate fluors with overlapping emission spectra. The user provides a signature spectrum of each individual fluor, and the instrument then mathematically separates the spectrum into the separate components. Triple images with cfp, gfp and yfp should be completely separated. This ability to mathematically separate overlapping fluors is unique to the Zeiss. The ability to record the full spectrum of each image point will be extremely powerful for Fluorescence Resonance Energy Transfer. Another specialty of the Zeiss is its "physiology" software package, which provides a very user-friendly facility for Fluorescence Recovery after Photobleaching. All of these capabilities are addressed in proposed projects. We also propose to purchase a module for Fluorescence Correlation Spectroscopy (FCS). This module illuminates a tiny volume and measures the fluctuations of fluorescently labeled molecules. This provides a means to assay for the diffusion coefficient, and thereby the polymer size, of labeled macromolecules. This can be applied to study molecules in solution, but more importantly, when used in conjunction with the scanning confocal mode, can perform these measurements in cells or tissues. Projects for this module are proposed by two faculty from Cell Biology and two from Biochemistry. This will be the only FCS facility at Duke University, and we expect additional projects from across the campus.

描述（由申请人提供）： 该应用程序是购买现代激光扫描共聚焦显微镜，主要满足杜克大学医学中心细胞生物学系的需求。目前，唯一的部门共享是10岁的Bio-Rad MRC 600，在几个领域的功能有限。我们选择的显微镜是新的Zeiss LSM 510 meta。元排放探测器提供了一个独特而强大的检测系统，该系统远远超过了常规的二分色滤波器。元检测器基于一个衍射光栅，将发射散布到全光谱中，而检测器则记录每个扫描点的全光谱。这提供了独特的功能，可以通过重叠的发射光谱分离荧光。用户提供了每个单独荧光的签名频谱，然后将仪器数学分离到单独的组件中。具有CFP，GFP和YFP的三重图像应完全分开。这种数学上分离重叠荧光的能力是蔡司所独有的。记录每个图像点的完整频谱的能力对于荧光共振能量传递将非常强大。蔡司的另一个专长是其“生理学”软件包，该软件包为光漂白后提供了非常用户友好的荧光恢复设施。所有这些功能都在拟议的项目中解决。我们还建议购买用于荧光相关光谱（FCS）的模块。该模块照亮了很小的体积，并测量了荧光标记的分子的波动。这提供了一种分析扩散系数的方法，从而分析了标记的大分子的聚合物大小。这可以应用于研究溶液中的分子，但更重要的是，当与扫描共聚焦模式结合使用时，可以在细胞或组织中执行这些测量。该模块的项目由细胞生物学的两个教师和两个来自生物化学的教师提出。这将是杜克大学唯一的FCS设施，我们希望整个校园的其他项目。