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）： 本申请是购买一台现代激光扫描共聚焦显微镜，主要用于满足杜克大学医学中心细胞生物学系的需要。目前唯一的部门共聚焦是一个10岁的Bio-Rad MRC 600，它在几个领域的能力有限。我们选择的显微镜是新的蔡司LSM 510 Meta。Meta发射探测器提供了一个独特而强大的探测系统，远远超过传统的二向色滤光片。Meta探测器基于衍射光栅和探测器，衍射光栅将发射扩展为全光谱，探测器记录每个扫描点的全光谱。这提供了分离具有重叠发射光谱的荧光体的独特能力。用户提供每个荧光体的特征光谱，然后仪器通过数学方法将光谱分离为单独的组分。三重图像与cfp，gfp和yfp应完全分开。这种数学上分离重叠荧光的能力是蔡司独有的。记录每个图像点的全光谱的能力对于荧光共振能量转移来说将是极其强大的。蔡司的另一个特色是它的“生理学”软件包，它为光漂白后的荧光恢复提供了一个非常用户友好的工具。所有这些能力都在拟议的项目中得到解决。我们还建议购买一个荧光相关光谱（FCS）模块。该模块照亮一个微小的体积，并测量荧光标记分子的波动。这提供了一种测定扩散系数的方法，从而测定标记的大分子的聚合物尺寸。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进行这些测量。该模块的项目由细胞生物学的两名教师和生物化学的两名教师提出。这将是杜克大学唯一的FCS设施，我们预计整个校园的其他项目。