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）购买一台集成TIRF/活细胞共聚焦/快速宽视场荧光显微镜，该显微镜将被安置在MCB/生物科学学院成像设施部分。该仪器提供了重要的新的活细胞成像能力，这是目前设施中现有的两种“交钥匙”仪器无法实现的。该仪器还将在现有的共聚焦点扫描上节省时间，这将在明年饱和。该仪器基于3i马里亚纳实时共聚焦SDC工作站，通过共享激光激励源（TIRF和多孔径共聚焦所需）和灵敏的EM-CCD相机构建。该系统配置有两个独立的发射端口，包含三种成像模式。一个用于TIRF和快速宽视场应用，另一个用于多光圈共聚焦。在TIRF/宽视场和共聚焦之间的变化是通过一个机动光纤开关来完成的，该开关很容易由用户操作。2006年12月，我们在3i的设施中进行了为期一周的演示，证明了这种仪器设计对所有三种成像模式都有效。MCB/CBS成像设施目前为大量的UC-D研究人员提供服务（2005-2006财政年度17个部门的64名研究人员）。这里要求的复杂和多功能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。该提案中的三个主要用户在我们的仪器演示中获得了重要的新结果，这些结果是设施中现有仪器无法获得的。为了认识到该仪器将对加州大学戴维斯分校的研究事业作出的重要贡献，大学行政部门已认捐107 691美元作为购买价格。拟议仪器的管理将纳入现有的成功管理计划（在2005-2006财政年度，仪器使用时数为2119小时，充值费用为96,402元）。该计划由一位博士级别的科学家监督，并由一个院系影像委员会指导。

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.