Laser Scanning Confocal Microscope/Multicolor Spectra

激光扫描共焦显微镜/多色光谱

• 批准号: 6580688
• 负责人: WILLIAM J BETZ
• 金额: $ 38.05万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6580688
• 关键词: biomedical equipment purchase; confocal scanning microscopy; light microscopy

DESCRIPTION (provided by applicant): We request funds to purchase a Zeiss LSM 510 Meta confocal laser scanning microscope. Seven Major Users and five Minor Users, well funded by NIH (e.g., Principal Investigators on 18 R01 s, 1 PPG, 2 Center Grants, 3 R32 Training grants), describe 18 research projects that span a broad range of biological questions and will collectively utilize all of the capabilities of the instrument. The applicants represent seven different departments in the School of Medicine. The Dean has made a commitment of $250,000 for support of the instrument, which will be housed in and managed by a core Light Microscopy Facility. Most Users have had extensive experience with various imaging techniques, and have acquired sound preliminary data that illustrate both the need for this equipment and the feasibility of the proposed experiments. A comprehensive inventory of existing equipment and careful scrutiny of proposals make clear that none of the projects can be accomplished with current equipment at the Health Sciences Center or any other campus in reasonable proximity. A stable and time-tested management plan that has emerged over the past 10 years of continuous administration of shared optical facilities by the same personnel will insure that the instrument will be well maintained, that it will become self-supporting, and that it will be used intensively by well-trained scientists drawn both for the User Group and, over time, from the larger population of scientists in the region. The specific proposals all relate to existing funded R01 research grants, but were not originally included as part of those applications because of the lack of equipment to perform them. Almost all projects involve studies of trafficking of molecules and organelles in living cells, using Green Fluorescent Protein and its analogs as markers. The most important attribute of the requested instrument is the ability to produce emission spectra through 'lambda stack' acquisition and 'linear unmixing' of multiple fluorophores, characteristics that were documented first hand by Users in pilot studies. Other projects involve tracking movements of molecules by Fluorescence Recovery After Photobleaching (FRAP), and protein-protein interactions with Fluorescence Resonance Energy Transfer (FRET). Extensive User experience with these techniques insures that the hypotheses put forward are testable, and that results will be reliable.

描述(由申请人提供)： 我们请求资金购买蔡司LSM 510Meta共聚焦激光扫描显微镜。由美国国立卫生研究院资助的7个主要用户和5个次要用户(例如，18 R01 S，1个PPG，2个中心资助，3个R32培训资助)描述了18个研究项目，这些项目涵盖了广泛的生物学问题，并将共同利用该仪器的所有能力。申请者来自医学院的七个不同部门。院长承诺为该仪器提供25万美元的支持，该仪器将安装在一个核心的光学显微镜设施中并由其管理。大多数用户对各种成像技术都有丰富的经验，并获得了可靠的初步数据，说明了对该设备的需求和拟议实验的可行性。对现有设备的全面清点和对提案的仔细审查清楚地表明，在健康科学中心或距离合理的任何其他校园，利用现有设备无法完成任何项目。在过去10年由同一人员持续管理共享光学设施的过程中，出现了一个稳定和久经考验的管理计划，该计划将确保仪器得到很好的维护，使其成为自给自足的仪器，并确保训练有素的科学家为用户组以及随着时间的推移，从该区域更多的科学家群体中抽出大量资金进行密集使用。 这些具体建议都涉及现有的R01研究资助，但最初并未作为这些申请的一部分，因为缺乏执行这些资助的设备。几乎所有的项目都涉及研究活细胞中分子和细胞器的运输，使用绿色荧光蛋白及其类似物作为标记。所要求的仪器最重要的属性是能够通过对多个荧光团的“波长叠加”采集和“线性分解”产生发射光谱，用户在试点研究中第一手记录了这些特征。其他项目包括通过光漂白后荧光恢复(FRAP)跟踪分子运动，以及利用荧光共振能量转移(FRET)跟踪蛋白质-蛋白质相互作用。这些技术的广泛用户经验确保了提出的假设是可检验的，结果将是可靠的。