Acquisition of a Carl Zeiss Cell Observer Spinning Disk Confocal Microscope

购买卡尔蔡司细胞观察转盘共焦显微镜

• 批准号: 8447687
• 负责人: J. Michael MCCAFFERY
• 金额: $ 53.56万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-05 至 2014-07-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8447687
• 关键词: 4D Imaging; Afferent Neurons; Arts; Biomedical Engineering; Carbon Dioxide; Cell Differentiation process; Cells; Cellular biology; Circadian Rhythms; Computer software; Development; Endocytosis; Engineering; Ensure; Fluorescence Resonance Energy Transfer; Funding; Gene Expression Regulation; Germ Cells; Herpesviridae; Humidity; Image; Imaging Techniques; Incubators; Internet; Laboratories; Lasers; Letters; Life; Maintenance; Membrane; Membrane Protein Traffic; Microscope; Microscopy; Microtubules; Mitochondria; Molecular; Motor; Photobleaching; Phototoxicity; Recovery; Research; Research Personnel; Resources; Schools; Science; Site; Speed; System; Temperature; Time; United States National Institutes of Health; Universities; Virus Assembly; Work; base; cellular imaging; fluorophore; interest; medical schools; migration; neuron development; neuronal circuitry; photoactivation

DESCRIPTION (provided by applicant): A diverse user-group of 12 investigators, within The Johns Hopkins University School of Arts & Sciences, Whiting School of Engineering, and the School of Medicine, request funds to purchase a Carl Zeiss Cell Observer SD. This system, based on the Carl Zeiss Axio Observer inverted research microscope, integrates a Yokogawa spinning disk confocal with: 1) 405nm/488nm/561nm/635nm lasers; 2) two ultra-sensitive Evolve EMCCD cameras for simultaneous two-channel imaging; 3) piezo-electric focusing for fast/precise Z-stack acquisition; and 4) a cage incubator for precise modulation of temperature, CO2, and humidity in extended live- imaging applications. All of these components are integrated, and conveniently accessed and precisely controlled through the Carl Zeiss AxioVision software. This new state-of-the-art workstation will be sited within the Hopkins' Integrated Imaging Center (IIC, http://www.jhu.edu/iic) on the Homewood campus; and constitute a significant upgrade to the live imaging capabilities for our Homewood/Hopkins-wide investigators whose needs have grown significantly over the previous decade. The IIC is a Homewood campus/Hopkins-wide microscopy resource, jointly supported by the Krieger School of Arts and Sciences and the Whiting School of Engineering (see Ball & Douglas letter, page 2 of the appendix); and utilized regularly by multiple schools and departments comprising >100 laboratories and >300 users. Our investigators, all well funded through the NIH, work on a host of diverse basic cell biology, biomolecular, and bioengineering related questions including, though not limited to: cell differentiation and gene regulation; Herpes virus assembly and maturation; mitochondrial fission; membrane organization and dynamics; microtubule-based motors; membrane trafficking; germ cell development and migration; sensory neuron development; molecular mechanisms of endocytosis; and circadian rhythm neuronal circuitry. In the requested configuration, the Cell Observer SD will afford our investigators the capability to precisely image multiple fluorophores simultaneously or sequentially at real-time acquisition speeds; and the ability to accurately modulate laser power to greatly minimize phototoxicity and photobleaching during extended time-lapse imaging sessions. Additionally, our investigators variously require the advanced live cell imaging techniques of FLIP, FRET, FRAP, and photoactivation of GFP, and 4D imaging. The new system will be incorporated into the IIC's existing, well established recharge system to ensure recovery of funds for supplies and maintenance; it will be made freely available to all interested users Hopkins- wide through our convenient web-scheduler; and it will be incorporated into the IIC's annual undergraduate/graduate course offerings. (http://www.jhu.edu/iic/academic.htm).

描述（由申请人提供）：约翰霍普金斯大学艺术与科学学院、怀廷工程学院和医学院的12名研究人员组成了一个多样化的用户组，要求资金购买卡尔蔡司细胞观察SD。该系统基于卡尔蔡司Axio观察者倒置研究显微镜，集成了一个横河旋转盘共聚焦：1)405nm/488nm/561nm/635nm激光器；2)两台超灵敏的Evolve EMCCD相机，实现双通道同步成像；3)压电聚焦，实现快速/精确的z堆叠采集；4)在扩展的实时成像应用中，用于精确调制温度、二氧化碳和湿度的笼式培养箱。所有这些组件都集成在一起，通过卡尔蔡司AxioVision软件方便地访问和精确控制。这个新的最先进的工作站将位于霍姆伍德校园的霍普金斯综合成像中心（IIC, http://www.jhu.edu/iic）内；为霍姆伍德/霍普金斯大学范围内的调查人员提供了实时成像能力的重大升级，他们的需求在过去十年中显著增长。IIC是霍姆伍德校园/霍普金斯大学范围内的显微镜资源，由克里格艺术与科学学院和怀廷工程学院共同支持（见Ball & Douglas信函，附录第2页）；被多个院系、实验室100余家，用户300余家定期使用。我们的研究人员都是由美国国立卫生研究院资助的，他们研究了许多不同的基本细胞生物学、生物分子和生物工程相关的问题，包括但不限于：细胞分化和基因调控；疱疹病毒的组装和成熟；线粒体分裂;膜组织与动力学；microtubule-based汽车;膜贩运;生殖细胞发育和迁移；感觉神经元发育；内吞作用的分子机制；以及昼夜节律神经回路。在要求的配置中，Cell Observer SD将为我们的研究人员提供以实时采集速度同时或顺序精确成像多个荧光团的能力；并且能够准确地调制激光功率，以极大地减少在长时间延时成像期间的光毒性和光漂白。此外，我们的研究人员还需要先进的活细胞成像技术，如FLIP、FRET、FRAP、GFP光激活和4D成像。新系统将纳入保险公司现有的完善的充值系统，以确保回收供应和维修的资金；它将通过我们方便的网络调度程序免费提供给所有感兴趣的用户；它将被纳入IIC的年度本科/研究生课程。(http://www.jhu.edu/iic/academic.htm)。