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）： 本申请旨在购买一台现代激光扫描共焦显微镜，主要满足杜克大学医学中心细胞生物学系的需求。目前唯一的部门共聚焦设备是使用了 10 年的 Bio-Rad MRC 600，其在多个领域的功能有限。我们选择的显微镜是新型蔡司 LSM 510 META。 META 发射检测器提供了独特而强大的检测系统，远远超过了传统的二向色滤光片。 META 探测器基于将发射扩展到全光谱的衍射光栅，以及记录每个扫描点的全光谱的探测器。这提供了分离具有重叠发射光谱的荧光体的独特能力。用户提供每种荧光体的特征光谱，然后仪器以数学方式将光谱分离成单独的成分。 cfp、gfp 和 yfp 的三重图像应完全分离。这种以数学方式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非常强大。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非常用户友好的设施。所有这些功能都在拟议的项目中得到解决。我们还建议购买荧光相关光谱 (FCS) 模块。该模块照亮微小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，我们预计整个校园还会有更多项目。