CONFOCAL MICROSCOPE FOR SHARED RESOURCES FACILITY

该子项目是利用由NIH/NCRR资助的共享仪器补助金提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是资助机构，不一定是研究者的机构。描述（由申请人提供）：本修订后的共享仪器资助申请申请资金用于购买徕卡TCS SP2 AOBS激光扫描共聚焦显微镜。该仪器将被整合到H. Lee Moffitt癌症中心和研究所的分析显微镜核心设备中。新的共聚焦系统将取代已有8年历史的老化共聚焦系统，该系统不能满足我们PHS资助的研究人员的需求。现有的共聚焦系统是一个老化的2-PMT / 2激光系统，不包含我们研究人员所需要的多色定量工作所需的激发和检测能力。拟议的徕卡共聚焦系统是现代的4-PMT系统，加上5个激光线源（近紫外405，蓝色，绿色，橙色和红色），为我们的7名小灵通资助的研究人员提供所需的资源，以推进他们的研究工作。具体而言，选择徕卡共聚焦的因素是AOBS和SP光谱检测器。徕卡AOBS系统采用无滤光片设计，与我们为本提案评估的其他共聚焦系统相比，可以产生更亮的图像。与其他使用光栅探测器的光谱共聚焦系统不同，AOBS SP光谱探测器使用棱镜来提高收集效率，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”功能是光谱分离能力，可以分离发射范围非常接近甚至重叠的荧光染料。同样，驱动该显微镜的软件被认为提供了三个共聚焦系统评估中最友好的用户界面。修订后的建议因支持这一文书的机构得到加强而得到加强。这台仪器的费用将由莫菲特癌症中心支付，包括每年25,000美元的服务合同，用于放置仪器的专用空间和分析显微镜核心设施的工资支持。此外，莫菲特癌症中心还承诺购买辅助项目，如活细胞成像设备、徕卡软件模块和Imaris Bitplane软件套件，用于3D共定位和体绘制分析。

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation Grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the grant, which is not necessarily the institution for the investigator. 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.