A Tri-modular TIRF/Live Cell Confocal/Fast Widefield Fluorescence Imaging System

描述(由申请人提供)：一个用户组，包括9名NIH资助的研究人员，申请498,113.89美元从智能成像创新公司(3i)购买集成的TIRF/活细胞共焦/快速宽视场荧光显微镜，安装在MCB/生物科学学院成像设施部。这台仪器提供了重要的新的活细胞成像能力，这是目前该设施中现有的两台“交钥匙”仪器所不可能实现的。这台仪器还将腾出时间在现有的点扫描共焦上，这将在明年饱和。该仪器以3i Marianas实时共焦SDC工作站为基础，共享激光激发源(TIRF和多口径共焦所需)和灵敏的EM-CCD相机。该系统配置有两个独立的发射端口，包括三种成像模式。一个用于TIRF和快速广域应用，另一个专用于多孔径共焦。TIRF/宽视场和共焦之间的转换是使用电动光纤开关完成的，用户可以轻松操作。2006年12月，我们在我们的3i工厂进行了为期一周的演示，证明该仪器设计适用于所有三种成像模式。MCB/CBS成像设备目前服务于大量的UC-D研究人员(2005-2006财政年度17个部门的研究人员)。这里要求的复杂和多功能的仪器满足了高级研究人员的需求，他们的研究问题和成像需求已经超出了我们现有仪器的能力。本提案中的三个主要用户在我们的仪器演示期间取得了重要的新成果，这是该基金现有仪器无法获得的。鉴于该仪器将对加州大学戴维斯分校的研究事业做出重要贡献，大学行政部门已承诺为购买价格提供107,691美元。拟议仪器的管理将被整合到现有的、成功的管理计划中(2005-2006财政年度，2119个用户仪器小时和96,402美元的充值)，该计划由一名博士级别的科学家监督，并由教务系成像委员会指导。

DESCRIPTION (provided by applicant): A user group including nine NIH funded investigators requests $498,113.89 to purchase an integrated TIRF/live cell confocal/fast widefield fluorescence microscope from Intelligent Imaging Innovations (3i) to be housed in the Section of MCB/College of Biological Sciences Imaging Facility. This instrument provides important new live cell imaging capabilities not possible with the two existing "turn key" instruments currently in the facility. This instrument will also free time on the existing point scanning confocal, which will be saturated in the next year. The proposed instrument builds from a base 3i Marianas Real Time Confocal SDC workstation by sharing the laser excitation sources (required for TIRF and multiple-aperture confocal) and sensitive EM-CCD cameras. The system is configured with two separate emission ports that encompass three imaging modalities. One is shared for the TIRF and rapid widefield applications and the other dedicated to the multiple-aperture confocal. Changing between TIRF/widefield and confocal is accomplished using a motorized fiber optic switch which is easily operated by the user. We have demonstrated that this instrument design works well for all three imaging modalities during a weeklong demonstration in our facility by 3i in December 2006. The MCB/CBS Imaging Facility currently serves a very large number of UC-D researchers (64 researchers in 17 departments in fiscal year 2005-2006). The sophisticated and versatile instrument requested here meets the needs of advanced researchers whose research problems and imaging needs have outgrown the capabilities of our existing instrumentation. Three of the major users in this proposal obtained significant new results during our instrument demonstrations that could not be obtained with the existing instrumentation in the Facility. In recognition of the important contribution this instrument would make to the UC-Davis research enterprise, the university administration has pledged $107,691 towards the purchase price. Management of the proposed instrument will be integrated into the existing, successful management scheme (2119 user instrument hours and $96,402 of recharges in fiscal year 2005-2006), which is supervised by a PhD-level scientist and guided by a faculty departmental imaging committee.