Advanced Multi-color Confocal and FRAP-SAC Microscope

先进的多色共焦和 FRAP-SAC 显微镜

• 批准号: 7792767
• 负责人: JERROLD R. TURNER
• 金额: $ 49.99万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-16 至 2011-07-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7792767
• 关键词: Arts; Cancer Center; Cell Count; Cells; Chicago; Color; Computer software; Controlled Environment; Core Facility; Data; Development; Devices; Digestive System Disorders; Equipment; Faculty; Fluorescence Resonance Energy Transfer; Fluorescent Probes; Funding; Image; Incubators; Investigation; Laboratories; Lasers; Life; Lighting; Location; Malignant Neoplasms; Microscope; Neurologic; Preparation; Program Research Project Grants; Protocols documentation; Research; Research Personnel; Sampling; Scanning; Speed; Staging; System; Time; Training Support; Universities; aged; cellular imaging; charge coupled device camera; disability; fluorophore; instrument; light microscopy; meetings; member; photoactivation; research study

DESCRIPTION (provided by applicant): We are requesting funds to purchase a Marianas Real Time Confocal SDC microscope equipped with 1) a Yokogawa CSUX Nipkow scanner; 2) a motorized spherical aberration correction (SAC) device; 3) five laser lines to enable use of a broad range of fluorophores, including those used for FRET; 4) high-speed laser illumination for bleaching (or photoactivation) studies of fluorescent targets; 5) a dual camera/ image splitter system with high speed Photometrics Evolve EM-CCD cameras; and 6) an enclosed CO2-regulated incubator. This confocal system is necessary to meet the immediate research needs of the major users and other faculty, as it provides the new features described above, particularly SAC, FRET, FRAP, and simultaneous two color imaging within a controlled environment. In addition, the Marianas Real Time Confocal SDC microscope will increase the capacity of available instruments, which is critical given the increasing numbers of live cell imaging studies. These studies often require large blocks of time that have been unavailable in an increasing number of cases, particularly after the decommissioning of our aged Fluoview system. The new instrument will be incorporated into the Integrated Light Microscopy Core (ILMC) Facility, which presently serves over 165 laboratories at the University of Chicago, as well as a large number of multiple collaborative users from neighboring campuses. Nearly all facility users are NIH-funded investigators with the majority members of the University of Chicago Cancer Center Program Project Grant. The bulk of research conducted using the new platform will directly support investigations related to cancer, digestive disease, and neurological development and disabilities. Apart from adding state-of-the-art, high-speed, two-color imaging capabilities, the platform will perform as an additional multi-color confocal platform for live samples, which require large blocks of equipment access time. The ability to simultaneously co-observe two or more fluorescent probes during high speed confocal imaging is a critical need that is not met by existing equipment due to limitations of hardware. The imaging system is optimized for live cell observations by inclusion of an incubator system and xy scanning stage, which will allow users to automatically image multiple locations in preparations during time-lapse studies, greatly increasing the data throughput by decreasing the need for repeating long experiments just to increase the number of cells observed. Importantly, the software is intuitive and provides utilities for storage of acquisition protocols and macros. The similarity of the software to other systems in the ILMC will permit easy transition of existing users to the new platform. Core staff will provide support and training of users to quickly embrace the technological advances enabled by the proposed system.

描述(由申请者提供)：我们申请资金购买Marianas实时共聚焦SDC显微镜，配备1)横河CSUX Nipkow扫描仪；2)机动球面像差校正(SAC)设备；3)五条激光线路，以能够使用广泛的荧光团，包括用于FRET的荧光团；4)用于荧光目标漂白(或光激活)研究的高速激光照明；5)双摄像机/图像分光器系统，配备高速光度测量进化EM-CCD摄像机；以及6)封闭式二氧化碳调节孵化器。该共焦系统是满足主要用户和其他教员的即时研究需求所必需的，因为它提供了上述新特征，特别是SAC、FRET、FRAP和受控环境中的同时双色成像。此外，Marianas实时共聚焦SDC显微镜将增加现有仪器的容量，鉴于活细胞成像研究的数量不断增加，这一点至关重要。这些研究通常需要大量的时间，在越来越多的情况下是无法获得的，特别是在我们陈旧的FluoView系统退役之后。新仪器将被纳入集成光学显微镜核心(ILMC)设施，该设施目前为芝加哥大学的165多个实验室以及来自邻近校园的大量多个协作用户提供服务。几乎所有的设施用户都是由NIH资助的研究人员，其中大多数是芝加哥大学癌症中心项目资助项目的成员。使用新平台进行的大部分研究将直接支持与癌症、消化系统疾病以及神经发育和残疾相关的调查。除了增加最先进的高速双色成像能力外，该平台还将作为一个额外的多色共焦平台用于现场样品，这需要大量的设备访问时间。在高速共聚焦成像过程中同时观察两个或多个荧光探针的能力是一项关键需求，由于硬件的限制，现有设备无法满足这一需求。通过包括孵化器系统和XY扫描平台，成像系统针对活细胞观察进行了优化，这将允许用户在延时研究期间自动对制剂中的多个位置进行成像，通过减少仅为增加观察细胞数量而重复长时间实验的需要，极大地提高了数据吞吐量。重要的是，该软件是直观的，并提供用于存储采集协议和宏的实用程序。该软件与ILMC中其他系统的相似性将使现有用户能够很容易地过渡到新平台。核心工作人员将为用户提供支持和培训，使其能够迅速接受拟议系统带来的技术进步。