Zeiss LSM 510 Laser Scanning Microscope

描述(由申请者提供)：由美国国立卫生研究院资助的9名高产人员 调查人员在两个国家卫生研究院资助的19个项目(&gt；3,200,000美元ADC) 科罗拉多州立大学各系申请341,154美元的NCRR资金 购买蔡司LSM 510共焦激光扫描显微镜，以取代 现已停业但之前曾被大量使用的10岁小萨拉斯特罗/分子动力学 多探头2001共聚焦显微镜。在过去的四年里，该部 解剖学和神经生物学已经花费了超过4.7万美元来保留Sarstro 显微镜工作正常，但它在关键实验中经常失败， 它的能力如此有限，以至于现在只对一个群体有用， 它只用它来获得低分辨率的组织地图 冻结断裂分析。因此，所有主要用户组的成员必须 要么前往其他城市获取数据，要么他们必须依赖 其他州和加拿大的合作者获取可发布的数据。扫描速率 对于蔡司共焦来说，速度至少是以前的20倍 仪器(每分钟156帧与最大每分钟8帧) 512×512)，从而能够分析活细胞中的瞬时现象。 本申请中配置的蔡司LSM 510有三个激光器 (不可分级到4)、自动和可重现的图像捕获；3 同时记录荧光通道(加上可见光)；以及 同时隔行扫描双激光扫描(“多重跟踪”，不适用于 大多数其他共焦系统)。后一种功能允许捕获图像 足够快，以允许实时分析生物过程 活细胞(NIH资助的至少6个项目所必需的)。同样重要的还有 作为一个多用户设施，LSM 510具有坚固的设计和直观的成像 具有预置模式的软件，可供常规用户和专家用户使用 (这消除了大多数手动调整)。共聚焦显微镜在 研究钙信号、膜分子组成的项目， 神经元传递的突触和非突触机制及其相互关系 膜分子组成的变化与生理的变化 活动和/或疾病状态。蔡司510将安装在数字 影像中心，其运营100%由卫生部资助 解剖学和神经生物学，允许使用设备，每小时免费(0美元) 用户。一名半职主任将维护仪器并培训新用户。 DIC董事会将每两个月举行一次会议，以确保持续有效率 更换共焦显微镜的时间安排和操作，以考虑 未来的升级，并建立或更改DIC访问策略。

DESCRIPTION (provided by applicant): Nine highly-productive NIH-funded investigators with nineteen NIH-funded projects (>$3,200,000 ADC) in two departments at Colorado State University request $341,154 in NCRR funds to purchase a Zeiss LSM 510 Confocal Laser Scanning Microscope to replace a now-defunct but previously heavily-used 10-year old Sarastro/Molecular Dynamics MultiProbe 2001 Confocal Microscope. In the past four years, the Department of Anatomy and Neurobiology has spent more than $47,000 to keep the Sarastro microscope functioning, but it has failed so often during critical experiments, and it is of such limited capability that it is now useful to only one group, which uses it solely to obtain low resolution maps of tissues for freeze-fracture analysis. Thus, members of all of the Major Users groups must either travel to other cities to obtain data, or they must rely on collaborators in other states and Canada to obtain publishable data. Scan rates for the Zeiss confocal are at least 20 times faster than the previous instrument (156 frames per minute vs. maximum of 8 frames per minute at 512x512), thereby permitting analysis of transient phenomena in living cells. The Zeiss LSM 510 as configured in this application has three lasers (ungradable to four), automated and reproducible image capture; three fluorescence channels (plus visible light) recorded simultaneously; and simultaneous interlaced dual laser scanning ("multi-tracking," not available on most other confocal systems). The latter capability allows image capture sufficiently rapidly to permit real-time analysis of biological processes in living cells (essential for at least 6 NIH-funded projects). Also important for a mult-iuser facility, the LSM 510 has a robust design and intuitive imaging software, with preset modes for both aboutroutine" and aboutexpert" users (which eliminates most manual adjustments). Confocal microscopy is essential in projects investigating calcium signaling, molecular composition of membranes, synaptic and non-synaptic mechanisms of neuronal transmission, and correlation of alterations in membrane molecularcomposition with changes in physiological activity and/or disease state. The Zeiss 510 will be installed in the Digital Imaging Center, whose operation is 100 percent funded by the Department of Anatomy and Neurobiology, allowing use of equipment at no ($0) hourly charge to users. A half-time Director will maintain the instrument and train new users. The DIC Board of Directors will meet bi-monthly to insure continued efficient scheduling and operation of the replacement confocal microscope, to consider future upgrades, and to establish or change DIC access policies.