Upgrade Existing Multiwell, Fluorescence Timelapse Imaging Workstation to Confoca

将现有多孔、荧光延时成像工作站升级至 Confoca

• 批准号: 7595619
• 负责人: SUSETTE C MUELLER
• 金额: $ 23.69万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-14 至 2010-05-13
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7595619
• 关键词: Academic Medical Centers; Bite; Cancer Center; Cancer Center Support Grant; Cells; Chimeric Proteins; Color; Comprehensive Cancer Center; Computer software; Development; Drug Delivery Systems; Fees; Fluorescence; Funding; Grant; Hour; Housing; Hypertension; Image; Immunity; Infection; Lasers; Life; Location; Microscope; Microscopy; Migration Assay; Modality; Paper; Preclinical Drug Evaluation; Preparation; Principal Investigator; Proteins; Publishing; Research; Research Personnel; Research Support; Resource Sharing; Screening procedure; Speed; Staging; Subcellular structure; System; Tissues; Training and Education; Tumor Cell Invasion; United States National Institutes of Health; angiogenesis; cell behavior; charge coupled device camera; fluorescence imaging; fluorescence microscope; instrument; instrumentation; neoplastic cell; oncology; professor; public health relevance; research study

DESCRIPTION (provided by applicant): Our original epifluorescence timelapse system was purchased in 2001 (Shared Instrumentation grant 1 S10 RR15768-01, "Fluorescence Imaging of Live Cells and Tissues", principal investigator: Mueller). It is critical need of upgrading to fulfill requirements for multicolor, multiwell, timelapse imaging for a core of nine NIH funded investigators. Although the Metamorph software and computer have been upgraded once using Lombardi Comprehensive Cancer Center Funds (LCCC), and a screening software module was purchased jointly by two Georgetown investigators in 2006 (Mueller and Elmendorf), this instrument is not confocal. The multiwell, automated imaging feature makes possible multiple, parallel experiments over hours or days and dramatically increases research efficiency and the power to compare treatment modalities under identical culture conditions. Past applications include cell migration assays, long term imaging to document changes in tumor cell behavior or in subcellular localization of GFP-chimeric proteins, and drug screening. The system consists of a Nikon Eclipse TE300 inverted epi-fluorescence microscope, ORCA-ER monochrome cooled CCD 12-bit camera, and MetaMorph image acquisition and analysis software version 7 with the Multiwell Screening and Analysis software module, a Sutter Instruments DG-4 high speed wavelength switcher for multi-color, live imaging, Prior ProScan motorized x,y,z stage controller, bright field and fluorescence shutters and filter wheels, and an environmental chamber. We request funds to convert this system to a confocal system with diode lasers, an EM-CCD camera for increased sensitivity, and a fast emission filter wheel. From 2004-2007, 18 investigators have utilized this instrument over 2230 hours. Four papers have been published, two submitted, and several are in preparation utilizing this microscope system. It is housed in the Microscopy and Imaging Shared Resource of the Lombardi Comprehensive Cancer Center (LCCC) which is directed by Dr. Mueller, a Professor of Oncology. The manager for this facility is Dr. Brian Koss. Drs. Xu and Govindan participate by directing the TIRF/ multiphoton component and education/ training, respectively. This facility is funded by LCCC, chargeback fees, and the LCCC Cancer Center Support Grant (2P30-CA-51008). PUBLIC HEALTH RELEVANCE: This proposal for funds to purchase a laser confocal upgrade to a research microscope will primarily support research at the Vincent T. Lombardi Cancer Center, a designated NIH Comprehensive Cancer Center and other investigators in the Georgetown University Medical Center and Main Campus. This laser confocal upgrade will facilitate the identification of cellular locations of proteins and subcellular structures that may cause tumor cell invasion and angiogenesis, development of hypertension, fungal and malarial infections and development of immunity, and effective drug targeting. This microscope will be placed in a pre-existing shared resource, used by a core of Lombardi investigators, and made available to other Georgetown investigators also supported by NIH-funded grants.

描述（由申请人提供）：我们的原始卵光荧光时间解体系统是在2001年购买的（共享仪器授予1 S10 RR15768-01，“活细胞和组织的荧光成像”，主要研究员：Mueller）。至关重要的需要升级，以满足多色，多井，时间元素成像的需求，以构成9位NIH资助的研究人员的核心。尽管使用Lombardi综合癌症中心基金（LCCC）（LCCC）升级了Metamorph软件和计算机，并且2006年乔治敦研究人员共同购买了筛选软件模块（Mueller和Elmendorf），但该仪器并不是共同点。多层自动成像功能使多个数小时或几天的多次平行实验可能会大大提高研究效率，并具有比较在相同培养条件下比较治疗方式的能力。过去的应用包括细胞迁移测定，长期成像以记录肿瘤细胞行为的变化或GFP-核蛋白的亚细胞定位以及药物筛查。该系统由尼康蚀Te300倒倒荧光显微镜，Orca-er单色冷却CCD 12位相机以及Metamorph Image获取和分析软件版本7，具有多井筛选和分析软件模块，Sutter Instrument荧光百叶窗和滤轮轮，以及环境室。我们要求资金将该系统转换为带有二极管激光器的共聚焦系统，EM-CCD摄像头增加灵敏度以及快速的发射过滤轮。从2004年至2007年，有18位调查人员在2230小时内利用了该工具。已经发表了四篇论文，两篇论文提交了，其中一些正在使用此显微镜系统进行准备。它位于伦巴第综合癌症中心（LCCC）的显微镜和成像共享资源中，该资源由肿瘤学教授Mueller博士指导。该设施的经理是Brian Koss博士。博士。 Xu和Govindan分别指导TIRF/ Multiphoton组件和教育/培训，参加。该设施由LCCC，退款费和LCCC癌症中心支持拨款（2P30-CA-51008）资助。公共卫生相关性：该资金购买激光共享升级到研究显微镜的提案将主要支持Vincent T. Lombardi Cancer Center，这是NIH NIH综合癌症中心和乔治敦大学医学中心和主要校园的其他研究人员。这种激光共焦升级将促进蛋白质和亚细胞结构的细胞位置的鉴定，这些细胞位置可能导致肿瘤细胞侵袭和血管生成，高血压的发育，真菌和疟疾感染以及免疫的发育以及有效的药物靶向。该显微镜将被放置在伦巴第研究人员核心使用的已有共享资源中，并提供给其他乔治敦研究人员，也得到了NIH资助的赠款的支持。