Multi-Function Laser Scanning Microscope

多功能激光扫描显微镜

• 批准号: 7595588
• 负责人: David W Piston
• 金额: $ 49.95万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7595588
• 关键词: Aging; Animal Model; Area; Biological; Biomedical Research; Cell Survival; Cells; Charge; Chemicals; Communities; Confocal Microscopy; Contract Services; Diabetes Mellitus; Dyes; Fluorescence Microscopy; Functional disorder; Funding; Image; Laboratories; Lasers; Life; Malignant Neoplasms; Microscope; Microscopy; Movement; Mus; Neoplasm Metastasis; Neurosciences; Neurotransmitters; Optics; Productivity; Protocols documentation; Qualifying; Quantum Dots; Reaction; Research; Research Infrastructure; Resolution; Resource Sharing; Resources; Sapphire; Scanning; Services; Spectrum Analysis; Stimulus; System; Techniques; Thick; Time; Tissues; Training; Training and Education; Universities; cellular imaging; detector; diabetic; in vivo; instrument; instrumentation; mutant; novel strategies; public health relevance; tumor growth; two-photon

DESCRIPTION (provided by applicant): We request funds to purchase a shared two-photon/confocal laser scanning microscope (Carl Zeiss LSM710 NLO), as a replacement for an aging Zeiss LSM510 system. The existing microscope was originally installed in 1998 and has been upgraded several times (for instance, adding the META detector in 2002), but we have now been informed that Carl Zeiss will discontinue service support for this older LSM510/Axiovert100 system in 2009. To assure experimental continuity, we will utilize much of our existing optics and accessory hardware on the new system, including the recently purchased Coherent Chameleon Ti:Sapphire laser. The current system has 6 major users focused in three broad research areas: cancer, diabetes, and neuroscience, and these users will constitute the major user group for the proposed instrument. All of these major users have qualifying NIH-funded projects that specifically include the use of two-photon excitation, so the guaranteed reliability of a system under service contract is crucial. As with the current instrument, the proposed instrument will be part of the Cell Imaging Shared Resource (CISR), of which Dr. Piston is the Scientific Director and Dr. Wells is the Managing Director. All major users will have access to the instrument and training through the established CISR infrastructure. Usage charges have supported the service contract for the existing LSM510 NLO over the last eight years, and we foresee no difficulty in continuing that arrangement for the LSM710. In addition, the CISR will also help identify and train new users of the two-photon instrument as their projects require. The Resource has an extensive track record of education, training, and productivity with over 300 lab groups at Vanderbilt University. Over the last 15 years, we have introduced shared access to confocal microscopy, live cell imaging, two-photon excitation, total internal reflection (TIRF) microscopy, fluorescence correlation spectroscopy, and deconvolution microscopy. These techniques all began with use by the more biophysical laboratories, but have become widely used by the general biomedical research community. For imaging of thick intact tissues or live animal models (such as the mouse), two-photon excitation is far superior to other approaches and permits high-resolution imaging at a level ~6 fold deeper than with confocal microscopy. The proposed instrument will be the only generally available two-photon excitation imaging system available at Vanderbilt University. Using two-photon excitation, we have developed in vivo quantitative laser scanning imaging protocols for NAD(P)H, Ca2+ indicator dyes, quantum dots, and GFP mutants, as well as photochemical activation protocols. Over the last few years, we have used these new approaches to elucidate many biological mechanisms involved in tumor growth and metastasis, diabetic pathophysiology, and neurotransmitter dynamics. Because of the imminent lack of service support for the current microscope, the requested instrumentation is critical to continue this research momentum. PUBLIC HEALTH RELEVANCE: Over the last decade, it has become apparent that the biological cell cannot be regarded as a "bag of chemicals" that carries out reactions. The exact three-dimensional arrangement of the cellular components is tremendously important, as are the time-dependent changes in this arrangement during the life of the cell and upon interaction with external stimuli. Recent advances in microscopy, such as the two-photon excitation microscope requested here, allow us to watch these arrangements and movements in living cells with minimal effects on cell viability.

描述（由申请人提供）：我们请求资金购买共享双光子/共焦激光扫描显微镜（Carl Zeiss LSM710 NLO），作为老化的 Zeiss LSM510 系统的替代品。现有的显微镜最初安装于 1998 年，并已进行了多次升级（例如，在 2002 年添加了 META 探测器），但我们现在获悉 Carl Zeiss 将在 2009 年停止对这款旧版 LSM510/Axiovert100 系统的服务支持。为了确保实验的连续性，我们将在新系统上利用大部分现有的光学器件和配件硬件，包括最近购买的 Coherent Chameleon Ti:Sapphire 激光器。目前的系统有 6 个主要用户，专注于三个广泛的研究领域：癌症、糖尿病和神经科学，这些用户将构成拟议仪器的主要用户组。所有这些主要用户都有合格的 NIH 资助项目，其中特别包括使用双光子激发，因此服务合同下系统的可靠性保证至关重要。与现有仪器一样，拟议的仪器将成为细胞成像共享资源 (CISR) 的一部分，Piston 博士是该资源的科学总监，Wells 博士是常务董事。所有主要用户都可以通过已建立的 CISR 基础设施使用该仪器并接受培训。过去八年中，使用费一直支持现有 LSM510 NLO 的服务合同，我们预计 LSM710 继续这种安排不会有困难。此外，CISR还将根据项目需要帮助识别和培训双光子仪器的新用户。该资源在范德堡大学 300 多个实验室小组的教育、培训和生产力方面拥有广泛的记录。在过去 15 年中，我们引入了共焦显微镜、活细胞成像、双光子激发、全内反射 (TIRF) 显微镜、荧光相关光谱和反卷积显微镜的共享访问。这些技术最初都是由更多的生物物理实验室使用，但已被一般生物医学研究界广泛使用。对于厚的完整组织或活体动物模型（例如小鼠）的成像，双光子激发远远优于其他方法，并且允许比共聚焦显微镜更深约 6 倍的水平进行高分辨率成像。拟议的仪器将是范德比尔特大学唯一通用的双光子激发成像系统。使用双光子激发，我们开发了 NAD(P)H、Ca2+ 指示剂染料、量子点和 GFP 突变体的体内定量激光扫描成像方案以及光化学激活方案。在过去的几年中，我们利用这些新方法阐明了涉及肿瘤生长和转移、糖尿病病理生理学和神经递质动力学的许多生物学机制。由于当前显微镜即将缺乏服务支持，所需的仪器对于继续这一研究势头至关重要。公共健康相关性：在过去的十年中，很明显，生物细胞不能被视为进行反应的“化学物质袋”。细胞成分的精确三维排列非常重要，这种排列在细胞生命周期中以及与外部刺激相互作用时随时间发生的变化也非常重要。显微镜技术的最新进展，例如此处要求的双光子激发显微镜，使我们能够观察活细胞中的这些排列和运动，同时对细胞活力的影响最小。