Acquisition of a Carl Zeiss Cell Observer Spinning Disk Confocal Microscope

描述（由申请人提供）：约翰霍普金斯大学文理学院、怀汀工程学院和医学院的12名研究人员组成的多样化用户组要求资金购买卡尔蔡司细胞观察员SD。该系统基于Carl Zeiss Axio Observer倒置研究显微镜，集成了Yokogawa旋转圆盘共聚焦：1）405 nm/488 nm/561 nm/635 nm激光器; 2）两个超灵敏Evolve EMCCD相机，用于同时双通道成像; 3）压电聚焦，用于快速/精确的Z堆叠采集;以及4）笼式培养箱，用于在延长的实时成像应用中精确调节温度、CO2和湿度。所有这些组件都集成在一起，通过Carl Zeiss AxioVision软件可以方便地访问和精确控制。这种新的最先进的工作站将设在霍普金斯的综合成像中心（IIC，http：//www.jhu.edu/iic）内的霍姆伍德校园;并构成了一个显着的升级，为我们的霍姆伍德/霍普金斯范围内的调查人员的实时成像能力，其需求已显着增长，在过去的十年。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汀工程学院共同支持（见Ball &道格拉斯信，附录第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，所有通过美国国立卫生研究院的资金，工作在一个主机的各种基础细胞生物学，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;疱疹病毒组装和成熟;线粒体分裂;膜组织和动力学;微管为基础的马达;膜运输;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;和昼夜节律神经元回路。在所要求的配置中，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;以及精确调制激光功率的能力，以在延长的延时成像会话期间最大限度地减少光毒性和光漂白。此外，我们的研究人员还需要FLIP、FRET、FRAP和GFP的光活化以及4D成像等先进的活细胞成像技术。新系统将纳入IIC现有的完善的充值系统，以确保回收供应和维护资金;它将通过我们方便的网络调度程序免费提供给霍普金斯范围内的所有感兴趣的用户;它将被纳入IIC的年度本科生/研究生课程。(http：//www.jhu.edu/iic/academic.htm）。

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).