Confocal Microscope

描述（由申请人提供）：国家的最先进的共聚焦显微镜是需要在爱荷华州州立大学（ISU）NIH资助的项目。ISU现有的多用户共焦系统由图像分析和共焦显微镜设施容纳和操作，但需要一个新的系统，因为：1）现有的Leica System TCS/NT（1998）老化，无法满足主要用户的活细胞成像需求，并且缺乏所需的附件，如FRET和3D重建，2）在100英里半径范围内，ISU的研究人员还没有其他具有这些能力的共焦系统。通过调查和讨论，我们已经确定Leica TCS SP 5将提供快速扫描速率，高分辨率，高灵敏度以减少光负载和对细胞的损伤，并能够微调激发和发射通道，以帮助同时采集NIH资助的五个主要项目以及小型项目所需的多个荧光通道，其中一些也是NIH资助的，在本提案中概述。这些研究人员在共聚焦显微镜方面有很多经验，但受到我们现有系统的限制。主要用户项目包括：轴突束形成、JIL-1激酶对染色质结构和基因表达的影响、PKC 4信号传导、氧化应激和神经元中的多巴胺能变性、用于评估基因表达和神经元干细胞发育和可塑性的适体标签以及反转录转座子基因靶向。国际空间大学的图像分析和共聚焦设施也非常适合为大小用户处理这一新系统。该设施由ISU生物技术办公室支持，自1984年以来一直存在。Ackermann（PI）和Carter（合作者）自1998年以来收购、操作和管理现有的共聚焦系统。Ackermann是该提案的次要用户，但有一个R 01资助的项目。该设施处理共焦系统的所有方面（维护，培训，使用，升级等）。自成立以来一直有一个活跃的用户委员会，并向生物技术办公室理事会提供年度报告。这种仪器与公共卫生有关，支持已经资助的NIH项目，从更好地了解基因表达到神经干细胞发育和可塑性。

DESCRIPTION (provided by applicant): State-of-the art confocal microscopy is required for NIH-funded projects at Iowa State University (ISU). The existing multi-user confocal system at ISU is housed and operated by the Image Analysis and Confocal Microscopy Facility, but a new system is needed, because: 1) the existing Leica System TCS/NT (1998) is aging, cannot handle the live cell imaging needs of major users, and lacks needed accessories such as FRET and 3D- reconstruction, and 2) no other confocal system with these capabilities is available to ISU investigators for a 100 mile radius. Through survey and discussion, we have determined that a Leica TCS SP5 would provide the fast scan rates, high resolution, high sensitivity to reduce light load and damage to cells, and ability to fine tune excitation and emission channels to aid in simultaneous acquisition of multiple fluorescence channels needed for five major NIH-funded projects as well as minor projects, some also NIH-funded, outlined in this proposal. These investigators have much experience in confocal microscopy, but are stifled by the limitations of our existing system. Major user projects include: axonal fascicle formation, JIL-1 kinase effects on chromatin structure and gene expression, PKC4 signaling, oxidative stress, and dopaminergic degeneration in neurons, aptamer tags for assessing gene expression and neuronal stem cell development and plasticity, and retrotransposon gene targeting. The Image Analysis and Confocal Facility at ISU is also well suited to handle this new system for the major and minor users. The facility is supported by the Office of Biotechnology at ISU, has existed since 1984. Ackermann (PI) with Carter (collaborator) acquired, operated, and managed the existing confocal system since 1998. Ackermann is a minor user in this proposal, but has an R01- funded project. The facility handles all aspects of the confocal system (maintenance, training, usage, upgrades, etc.) and has had an active user committee since inception and provides annual reports to the Council of the Office of Biotechnology. This instrumentation is relevant to public health in terms of supporting already funded NIH- projects that range from a better understanding of gene expression to neuronal stem cell development and plasticity.