Nikon A1R confocal laser microscope system for live cell imaging

描述（由申请人提供）：本提案的短期和长期目标是升级和扩大牙科医学（SDM）成像和分析核心设施的学校，使其可以继续与国家的最先进的设备和支持资助的研究人员谁需要活细胞成像能力的目标。该设施目前拥有SDM唯一的核心共聚焦显微镜，一台9年的BioRad Radiance 2000（R）。我们最近被告知，蔡司将不再提供这些仪器的服务合同（截至2009年9月），维修将只在零件可用时进行。因此，需要资金购买活细胞共聚焦成像（LCCI）系统，特别是尼康A1 R。我们选择了这个单光子高分辨率共焦系统，因为它允许慢和快扫描速度。为了提供从近UV（dapi/hoechst）到远红光（CY 5）的荧光团的激发和检测，该系统配备有405 nm二极管激光器、多线氩离子激光器、638二极管激光器和561 nm固态激光器。该系统的配置将有4个光电倍增管，用于采集多达4种颜色，以及一个32通道光谱检测器。选择此配置是为了提供灵活性，以满足我们用户组的各种需求。此外，它将支持我们的研究人员将他们的研究扩展到更大数量的荧光染料，从而获得每个细胞更多的信息。研究人员将能够监测：广泛的表面和细胞内标志物，评估细胞离子浓度（如钙）的变化，跟踪溶酶体成熟，分析病毒融合事件和信号传导途径，以及研究体内蛋白质-蛋白质和蛋白质-脂质相互作用。 核心研究设施对诸如可持续发展机制这样的研究企业的成功至关重要，因为核心研究设施向研究人员提供了在其研究活动中获得先进技术的机会，否则，由于财政原因，科学家个人或实验室将负担不起。SDM-IAC支持超过15名科学家的不同需求，他们的研究工作集中在促进口腔健康和了解疾病的基本机制。支持的项目特别侧重于研究口腔和颅面组织对生理和病理刺激的反应，重点是三个领域：炎症和免疫，传染病以及软硬结缔组织的生物学。

DESCRIPTION (provided by applicant): The short and long-term objective of this proposal is to upgrade and expand the School of Dental Medicine (SDM) Imaging and Analytical Core Facility so that it may continue to operate with state-of-the-art equipment and support the aims of funded investigators who require live cell imaging capabilities. This facility presently houses the only core confocal microscope at the SDM, a 9 year old BioRad Radiance 2000(R). We were recently informed that Zeiss will no longer provide service contracts for these instruments (as of Sept, 2009) and repairs will be made only when and if parts are available. Therefore funds are requested to purchase a live cell confocal imaging (LCCI) system, specifically a Nikon A1R. We chose this single photon high resolution confocal system with hydrid scanning capabilities because it allows for slow and fast scan speeds. In order to provide excitation and detection of fluorophores from the near UV (dapi/hoechst) to the far red (CY5) the system is equipped with a 405nm diode laser, a multiline Argon ion laser, a 638 diode laser and a 561nm solid state laser. The configuration of the system will have 4 PMTS for acquisition of up to 4 colors as well as a 32 channel spectral detector. This configuration was chosen to provide flexibility to meet the diverse needs of our user group. Moreover, it will support our investigators as they expand their studies to larger numbers of fluorochromes and thereby obtain significantly more information per cell. Investigators will be able to monitor: a wide range of both surface and intracellular markers, assess changes in cellular concentrations of ions such as calcium, follow lysosome maturation, analyze viral fusion events and signaling pathways as well as study protein-protein and protein-lipid interactions over time in vivo. Core research facilities are vital to the success of research enterprises such as that at the SDM by providing investigators with access to advanced technologies in their research activities, which, for financial reasons, would otherwise not be affordable to the individual scientist or laboratory. The SDM-IAC supports the diverse needs of over 15 scientists whose research efforts are focused on advancing oral health and understanding basic mechanisms of disease. Supported projects are focused, in particular, on studies on the response of oral and craniofacial tissue to physiologic and pathologic stimuli with emphasis in three areas: inflammation and immunity, infectious disease, and the biology of soft and hard connective tissue.