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）：我们最初的落射荧光延时系统于 2001 年购买（共享仪器拨款 1 S10 RR15768-01，“活细胞和组织的荧光成像”，主要研究员：Mueller）。迫切需要升级以满足 NIH 资助的九名核心研究人员对多色、多孔、延时成像的要求。尽管 Metamorph 软件和计算机已经使用 Lombardi 综合癌症中心基金 (LCCC) 进行了升级，并且筛查软件模块是由乔治敦大学的两名研究人员（Mueller 和 Elmendorf）于 2006 年联合购买的，但该仪器不是共聚焦的。多孔自动成像功能使得在数小时或数天内进行多个并行实验成为可能，并显着提高了研究效率以及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、记录肿瘤细胞行为或 GFP 嵌合蛋白亚细胞定位变化的长期成像以及药物筛选。该系统由 Nikon Eclipse TE300 倒置落射荧光显微镜、ORCA-ER 单色冷却 CCD 12 位相机以及带有 Multiwell 筛选和分析软件模块的 MetaMorph 图像采集和分析软件版本 7（Sutter Instruments DG-4 high）组成。用于多色、实时成像的高速波长切换器、Prior ProScan 电动 x、y、z 平台控制器、明场和荧光快门和滤光轮以及环境室。我们请求资金将该系统转换为带有二极管激光器的共焦系统、用于提高灵敏度的 EM-CCD 相机和快速发射滤光轮。从2004年到2007年，18名研究人员使用该仪器超过2230小时。已发表四篇论文，两篇已提交，还有几篇正在利用该显微镜系统准备。它位于隆巴迪综合癌症中心 (LCCC) 的显微镜和成像共享资源中，该中心由肿瘤学教授 Mueller 博士领导。该设施的经理是 Brian Koss 博士。博士。 Xu 和 Govindan 分别参与指导 TIRF/多光子组件和教育/培训。该设施由 LCCC、退款费和 LCCC 癌症中心支持补助金 (2P30-CA-51008) 资助。公共健康相关性：这项购买激光共焦升级研究显微镜的资金提案将主要支持 Vincent T. Lombardi 癌症中心、指定的 NIH 综合癌症中心以及乔治城大学医学中心和主校区的其他研究人员的研究。这种激光共聚焦升级将有助于识别蛋白质和亚细胞结构的细胞位置，这些位置可能导致肿瘤细胞侵袭和血管生成、高血压、真菌和疟疾感染的发展、免疫力的发展以及有效的药物靶向。该显微镜将放置在一个预先存在的共享资源中，由 Lombardi 的核心研究人员使用，并提供给其他乔治城研究人员使用，这些研究人员也得到了 NIH 资助的资助。