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，惠廷工程学院和医学院内的12位调查人员的多样化用户群，请求资金购买卡尔·蔡司牢房观察者SD。该系统基于Carl Zeiss Axio观察者倒立的研究显微镜，将横向川旋转磁盘共同集成了：1）405nm/488nm/561nm/561nm/635nm激光器； 2）两个超敏感的进化EMCCD相机，用于同时进行两通道成像； 3）对快速/精确Z堆栈采集的压电电气聚焦； 4）在扩展的活成像应用中，用于精确调节温度，二氧化碳和湿度​​的笼孵化器。所有这些组件都是集成的，并通过Carl Zeiss Axiovision软件进行了方便访问和精确控制。这个新的最先进的工作站将位于霍普金斯集成成像中心（IIC，http://www.jhu.edu/iic/iic）中；并为我们的Homewood/Hopkins范围的调查人员的实时成像功能构成了重大升级，这些调查员在过去的十年中的需求已经显着增长。 IIC是一家Homewood校园/霍普金斯范围内的显微镜资源，由克里格艺术与科学学院和工程惠廷学院共同支持（请参阅附录的第2页Ball＆Douglas Letter，第2页）；并由多个学校和部门定期使用，包括100个实验室和> 300个用户。我们的研究人员都通过NIH提供了充分的资金，从事许多不同的基本细胞生物学，生物分子和生物工程相关问题，包括但不限于：细胞分化和基因调节；疱疹病毒组装和成熟；线粒体裂变；膜组织和动态；基于微管的电动机；膜贩运；生殖细胞发育和迁移；感觉神经元发展；内吞作用的分子机制；和昼夜节律神经元电路。在请求的配置中，细胞观察者SD将使我们的研究人员能够在实时采集速度下同时或依次同时或依次地对多个荧光团进行多种图像；并能够准确调节激光功率以极大地最大程度地减少延长的延时成像会议期间的光毒性和光漂白。此外，我们的研究人员需要各种各样的flip，fret，frap和GFP光活化和4D成像的活细胞成像技术。新系统将纳入IIC现有的，良好的充值系统中，以确保恢复用于供应和维护的资金；它将通过我们方便的网络播种机免费提供给所有感兴趣的用户Hopkins- hopkins-wide；它将纳入IIC的年度本科/研究生课程。 （http://www.jhu.edu/iic/academic.htm）。