Nikon A1R confocal laser microscope system for live cell imaging

用于活细胞成像的 Nikon A1R 共焦激光显微镜系统

• 批准号: 7791744
• 负责人: Kathleen Boesze-Battaglia
• 金额: $ 49.99万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-27 至 2012-05-26
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7791744
• 关键词: 9 year old; Area; Argon; Arts; Biology; Calcium; Cells; Color; Communicable Diseases; Connective Tissue; Contract Services; Core Facility; Dental Schools; Detection; Disease; Equipment; Event; Fluorochrome; Funding; Housing; Image; Immunity; Individual; Inflammation; Ions; Laboratories; Lasers; Life; Lipids; Lysosomes; Medicine; Microscope; Monitor; Oral; Oral health; Organism; Pathologic; Photons; Physiological; Proteins; Research; Research Activity; Research Personnel; Resolution; Scanning; Scientist; Signal Pathway; Speed; Stimulus; Surface; System; Technology; Time; Tissues; Viral; cellular imaging; craniofacial; cyanine dye 5; detector; flexibility; fluorophore; in vivo; instrument; meetings; pyridinium 3-methoxyestra-1,3,5(10)-trien-6-yl sulfate; repaired; research facility; response; solid state; success

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.

描述(由申请人提供)：这项建议的短期和长期目标是升级和扩大牙科医学院(SDM)成像和分析核心设施，以便它可以继续使用最先进的设备，并支持需要活细胞成像能力的受资助研究人员的目标。该设施目前是SDM唯一的核心共聚焦显微镜，是一台已有9年历史的BioRad Radiance 2000(R)。我们最近接到通知，蔡司将不再为这些仪器提供服务合同(从2009年9月起)，只有在有部件可用时才会进行维修。因此，需要资金购买活细胞共聚焦成像(LCCI)系统，特别是尼康A1R。我们选择了这种具有混合扫描能力的单光子高分辨率共聚焦系统，因为它允许慢速和快速扫描。为了提供从近紫外光(DAPI/Hoechst)到远红光(Cy5)的荧光团的激发和检测，该系统配备了405 nm半导体激光器、多线氩离子激光器、638半导体激光器和561 nm固体激光器。该系统的配置将有4个PMT，用于采集多达4种颜色，以及一个32通道光谱探测器。选择这种配置是为了提供灵活性，以满足我们用户群体的不同需求。此外，它将支持我们的研究人员将他们的研究扩展到更多的荧光染料，从而获得更多的每个细胞的信息。研究人员将能够监测广泛的表面和细胞内标志物，评估钙等离子的细胞浓度变化，跟踪溶酶体成熟，分析病毒融合事件和信号通路，以及研究体内随着时间的推移蛋白质与蛋白质和蛋白质与脂质的相互作用。核心研究设施为研究人员提供在其研究活动中获得先进技术的机会，因此核心研究设施对研究企业的成功至关重要，例如在可持续发展研究中，由于资金原因，科学家或实验室个人负担不起这些技术。SDM-IAC支持超过15名科学家的不同需求，他们的研究重点是促进口腔健康和了解疾病的基本机制。所支持的项目特别侧重研究口腔和头面部组织对生理和病理刺激的反应，重点放在三个领域：炎症和免疫、传染病和软硬结缔组织生物学。