Acquisition of a Zeiss LSM 7 MP multiphoton microscope

描述(由申请人提供)：需要资金购买蔡司LSM MP多光子显微镜，配备相干变色龙超II钛：蓝宝石脉冲激光器。这台仪器将取代我们目前的BioRad Radiance多光子显微镜，后者不再满足我们用户的需求。此外，卡尔·蔡司正在逐步淘汰他们的传统BioRad系统，不再为Radiance 2300系统储备替换硬件或提供软件升级。专用多光子显微镜将安装在NCRR生物医学研究卓越中心(COBRE)神经科学拨款支持的成像/生理学核心设施中(5P20 RR16435，07/01/06-06/30/11)。科布雷赠款的竞争性续签预计将于2011年7月1日开始，并提供额外5年的核心支持。神经科学科布雷成像/生理学核心由一名全职、训练有素的显微镜专家管理。收购蔡司LSM 7 MP专用多光子显微镜，这是一个现代化、模块化和积极支持的系统，将极大地增强核心的能力，并将满足目前核心研究人员的新需求，并为更多的资助研究人员提供机会，显著扩大他们的研究范围 程序。新型蔡司多光子显微镜LSM 7MP具有惊人的快速采集速度，比我们的BioRad Radiance 2100 MPD双光子显微镜的速度快约4倍。LSM 7 MP的高速采集速度与卓越的灵敏度和较低的噪声水平相结合，使其成为研究大脑中活神经元和其他细胞类型活动的完美选择。近年来，笼状化合物的多光子光解(UNCAGING)被越来越多地作为一种工具，以极高的时间、位置和体积精确地将钙等物质快速输送到细胞内或细胞外空间。此外，与单光子光解相比，多光子去功能化可以更深入地渗透到活组织中，体积更小。我们高度创新的研究项目，研究大脑和脑小动脉平滑肌和内皮功能中的神经-血管耦合，绝对需要使用笼子里的钙、IP3、谷氨酸和ATP的光解。蔡司LSM 7 MP能够在不减慢采集速度的情况下，在多个任意定义的感兴趣区域内进行双光子光解。这将使我们能够同时对活细胞或组织进行去核和成像，其速度足以监测和记录因去核而发生的细胞活动的变化。此外，这将使我们能够同时或连续地将化合物连接到不止一个细胞或位置，从而为我们提供了研究大脑中不同细胞或结构之间的相互作用和通信的独特机会。 公共卫生相关性：该工具将支持旨在了解大脑和自主器官的发育和正常功能以及它们在应对疾病时的变化的研究。脑片和器官内细胞反应的成像将提供对神经元、血管和结缔组织功能的调节及其相互依赖的关键洞察。这一洞察力将为开发攻击这些组织的疾病的治疗方法提供信息。

DESCRIPTION (provided by applicant): Funds are requested to purchase a Zeiss LSM MP multiphoton microscope with a Coherent Chameleon Ultra II Ti:Sapphire pulsed laser. This instrument will replace our current BioRad Radiance Multiphoton Microscope, which no longer meets the needs of our users. In addition, Carl Zeiss is phasing out their legacy BioRad systems and no longer stocks replacement hardware or provides software upgrades for the Radiance 2300 system. The dedicated multiphoton microscope will be housed in the Imaging/Physiology Core Facility supported by the NCRR Center of Biomedical Research Excellence (COBRE) in Neuroscience grant (5P20 RR16435, 07/01/06 - 06/30/11). The competitive renewal of the COBRE grant is expected to start 7/1/01/2011 and provide 5 additional years of Core support. The Neuroscience COBRE Imaging/Physiology Core is administered by a full-time, highly trained microscopist. Acquisition of the Zeiss LSM 7 MP dedicated multiphoton microscope, a modern, modular, and actively supported system, would greatly enhance the capabilities of the Core and would meet the emerging needs of the present Core investigators and provide the opportunity for additional funded investigators to significantly expand the scope of their research programs. The new Zeiss multiphoton microscope, model LSM 7MP, offers impressively rapid acquisition speed, about 4 times higher than the speed of our BioRad Radiance 2100 MPD two-photon microscope. The high acquisition speed combined with the superior sensitivity and lower noise levels makes LSM 7 MP perfect for studying the activity of living neurons and other cell types in the brain. In recent years, multiphoton photolysis of caged compounds (uncaging) is increasingly used as a tool to quickly deliver calcium and other substances intracellularly or in the extracellular space with great precision in time, location and volume. Furthermore, multiphoton uncaging allows deeper penetration into live tissues, with smaller volumes compared to single photon photolysis. Our highly innovative research projects that investigate the neuro-vascular coupling in the brain and cerebral arteriole smooth muscle and endothelium function absolutely require the use of photolysis of caged calcium, IP3, glutamate and ATP. The Zeiss LSM 7 MP is capable of two photon photolysis localized within multiple arbitrarily defined regions of interest without slowing down the acquisition speed. This will allow us to simultaneously uncage and image living cells or tissue with a speed that is adequate for monitoring and recording the changes in cellular activity that occur as a result of uncaging. Moreover, this will enable us to simultaneously or consecutively uncage compounds in more than one cell or location thus providing us with the unique opportunity to study the interactions and communications between different cells or structures in the brain. PUBLIC HEALTH RELEVANCE: This instrument would support research designed to understand the development and normal functioning of the brain and autonomic organs and their alteration in response to disease. Imaging of cellular responses within brain slices and organs will provide key insight into the regulation of neuronal, vascular, and connective tissue function and their interdependence. This insight will inform development of therapeutic approaches for diseases that strike these tissues.