Confocal Microscope for Shared Resources Facility

描述（由申请人提供）：此修订后的共享仪器资助申请要求将资金用于购买Leica TCS SP2 AOBS激光扫描共聚焦显微镜。该仪器将被集成到H.李莫菲特癌症中心和研究所分析显微镜核心设施。新的共聚焦将取代8岁的老化共聚焦系统，不符合我们的PHS资助的研究者的需求。现有的共聚焦是一个老化的2-PMT / 2-激光系统，不包含我们的研究人员所需的定量工作所需的激发和检测能力。建议的徕卡共聚焦系统是现代的4-PMT系统，与5个激光线源（近紫外405，蓝色，绿色，橙子和红色）相结合，为我们的7个PHS资助的研究人员提供所需的资源，以推进他们的研究工作。选择Leica共聚焦的具体因素是AOBS和SP光谱检测器。Leica AOBS系统采用无滤光片设计，与我们评估的其他共焦系统相比，可产生更明亮的图像。与其他使用光栅探测器的光谱共焦系统不同，AOBS SP光谱探测器使用棱镜，以获得更高的收集效率，从而允许用户自由定义检测范围。该系统将为共享资源设施带来的另一个“增值”功能是光谱分解能力，可以分离发射范围非常接近甚至重叠的荧光染料。类似地，驱动该显微镜的软件被认为提供了所评估的三个共聚焦系统中最用户友好的界面。通过加强支持这一文书的机构，修订后的提案得到了加强。该仪器的成本将由莫菲特癌症中心购买每年25，000美元的服务合同，一个专门的空间来容纳仪器和分析显微镜核心设施的工资支持。此外，Moffitt Cancer还承诺购买辅助项目，如活细胞成像设备、Leica附加软件模块集合以及用于3D共定位和体绘制分析的Imaris Bitplane Suite软件。

DESCRIPTION (provided by applicant): This revised Shared Instrumentation Grant application requests funds to be used in the purchase of a Leica TCS SP2 AOBS Laser Scanning Confocal Microscope. This instrument will be integrated into the H. Lee Moffitt Cancer Center & Research Institute Analytic Microscopy Core Facility. The new confocal will replace an 8-year old aging confocal system that does not meet the needs of our PHS funded investigators. The existing confocal is an aging 2-PMT / 2-Laser system that does not contain the need excitation and detection capabilities to required for multicolor quantitative work required by our investigators. The proposed Leica confocal system is modern 4-PMT system coupled with 5 laser lines sources (Near UV 405, Blue, Green, Orange and Red) to give our 7 PHS funded investigators the needed resources to advance their research efforts. Specific, factors in selecting the Leica confocal were the AOBS and the SP spectral detector. The Leica AOBS system utilizes a filterless design to produce brighter images when compared to other confocal systems we evaluated for this proposal. Unlike other spectral confocal systems that use a grating based detector, the AOBS SP spectral detector uses a prism for greater collection efficiency allowing detection ranges to be freely defined by the user. Another "value added" feature that system will bring to the shared resourced facility are the spectral unmixing capabilities that can separate fluorochromes with very close or even overlapping emission ranges. Similarly the software driving this microscope was deemed as providing the most user friendly interface of the three confocal systems evaluated. The revised proposal is enhanced by a strengthened institutional supporting this instrument. The cost of this instrument will be matched by Moffitt Cancer Center with the purchase of a yearly $25,000 service contract, a dedicated space to house the instrument and salary support for Analytic Microscopy Core Facility. In addition, the Moffitt Cancer is committing to purchasing ancillary items such as live cell imaging equipment, collection of Leica add on software modules and the Imaris Bitplane Suite of software for 3D colocalization and volume rendering analysis.