Nipkow-disk Confocal Microscope for Live-Cell Imaging

描述（由申请人提供）：本申请支持购买Perkin-Elmer Ultraview LCI共聚焦显微镜。这种新设计的显微镜，基于最近在Nipkow磁盘技术的改进，代表了活细胞共聚焦成像的重大进步。与使用激光扫描的显微镜相比，Ultraview可以实现更快的图像采集，实时共聚焦观察，并且大大降低了光毒性（与单光子共聚焦显微镜相比）。指定的用户组由13名科学家组成，他们在NIH 8个不同研究所资助的27个研究项目中担任pi。关键项目需要在多个焦平面上用两个或多个荧光团标记的大区域上捕获共聚焦图像的时间序列，以便以m/秒的速度可视化运动。这些能力是一些参与研究人员的研究项目不可或缺的一部分，并将极大地有利于所有指定用户的研究工作。项目包括果蝇胚胎中的RNA转运、培养细胞中运动蛋白介导的运动、培养神经元的后高尔基分选和轴突转运、毛细胞中的pH变化和蛋白质运动、脑切片中的钙动力学、上皮单层中铁转运蛋白的运输、成纤维细胞中的脂筏相关离子通道和胰岛细胞中的钾通道。激光扫描显微镜还不能满足这些要求。所要求的仪器将配备附件，以便各种调查人员灵活使用。它将被安置在OHSU的活细胞成像设施中，该设施目前为35名注册用户提供服务，这些用户来自位于OHSU主校区的bb200个研究实验室。仪器的使用将由Bruce Schnapp（细胞和发育生物学教授）、Gary Banker （CROET高级科学家）和一个来自其他部门的指导委员会监督。Schnapp和Banker在先进成像方法方面拥有超过30年的专业知识。用户培训和仪器的日常操作将由Stefanie Kaech博士负责，她拥有10年的共聚焦成像经验，并在弗里德里希·米歇尔研究所（巴塞尔）工作期间参与了Ultraview的beta测试。

DESCRIPTION (provided by applicant): This application supports the purchase of a Perkin-Elmer Ultraview LCI confocal microscope. This newly designed microscope, based on a recent enhancement in Nipkow disk technology, represents a major advance in confocal imaging of living cells. Compared with microscopes that use laser scanning, the Ultraview enables far faster image acquisition, real-time confocal viewing, and greatly reduced phototoxicity (compared with single-photon confocals). The designated user group consists of 13 scientists who serve as PIs on 27 research projects funded by 8 different institutes of the NIH. Key projects demand capturing time-series of confocal images over large regions of interest in multiple focal planes labeled with two or more fluorophores in order to visualize movements at m/sec speeds. These capabilities are integral to the research programs of several of the participating investigators and will greatly benefit the research efforts of all designated users. Projects include studies of RNA transport in Drosophila embryos, kinesin-mediated movements in cultured cells, post-Golgi sorting and axonal transport in cultured neurons, pH changes and protein movements in hair cells, calcium dynamics in brain slices, trafficking of iron transport proteins in epithelial monolayers, lipid raft-associated ion channels in fibroblasts, and potassium channels in pancreatic islet cells. Laser scanning microscopes cannot begin to meet these demands. The requested instrument will be equipped with accessories to permit flexible use by a diverse group of investigators. It will be housed in OHSU's Live-Cell Imaging Facility, which currently serves a group of 35 registered users drawn from among the >200 research laboratories located on the main campus of OHSU. Use of the instrument will be overseen by Bruce Schnapp (Professor, Cell and Developmental Biology), Gary Banker (Senior Scientist, CROET), and a steering committee drawn from other departments. Together, Schnapp and Banker have more than 30 years of expertise with advanced imaging methods. User training and day-to-day operation of the instrument will be the responsibility of Dr. Stefanie Kaech, who has 10 years experience with confocal imaging and participated in beta-testing of the Ultraview while on the staff at the Friedrich Miescher Institute (Basel).