Laser Scanning Confocal Microscope/Multicolor Spectra

描述（由申请人提供）： 我们申请资金购买蔡司LSM 510 Meta共聚焦激光扫描显微镜。七个主要用户和五个次要用户，由NIH资助（例如，18个R 01、1个PPG、2个中心赠款、3个R32培训赠款的主要研究者描述了18个研究项目，这些项目涵盖了广泛的生物学问题，并将共同利用仪器的所有功能。申请人代表医学院的七个不同部门。院长已承诺为该仪器提供25万美元的支持，该仪器将被安置在一个核心光学显微镜设施中并由其管理。大多数用户都有各种成像技术的丰富经验，并已获得良好的初步数据，说明需要这种设备和拟议的实验的可行性。对现有设备的全面清点和对提案的仔细审查清楚地表明，没有一个项目可以用健康科学中心或任何其他合理附近的校园的现有设备来完成。在过去10年中出现了一个稳定和经过时间考验的管理计划，由相同的人员持续管理共用光学设施，这将确保仪器得到良好的维护，使其能够自我维持，并由训练有素的科学家集中使用，这些科学家既是为用户组挑选的，也是随着时间的推移从该地区更多的科学家中挑选的。 具体的建议都涉及现有的资助R 01研究赠款，但最初没有包括在这些应用程序的一部分，因为缺乏设备来执行它们。几乎所有的项目都涉及使用绿色荧光蛋白及其类似物作为标记物研究活细胞中分子和细胞器的运输。所需仪器的最重要属性是能够通过“λ叠加”采集和多个荧光团的“线性解混”产生发射光谱，用户在试点研究中直接记录了这些特征。其他项目包括通过光漂白后荧光恢复（FRAP）跟踪分子运动，以及通过荧光共振能量转移（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测试的，结果是可靠的。

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.