Building Enhanced Capability for the PennVet Multiphoton Core

 描述(由申请者提供)：为了应对日益增长的对先进的活细胞成像能力的需求，宾夕法尼亚大学兽医学院(SOVM)于2008年夏天建立了成像核心设施。在几个先驱实验室的参与下，以及大学普罗沃斯特办公室和SOVM中的部门和中心捐赠的近90万美元，我们购买了几台显微镜，包括徕卡SP5多光子系统。我们的核心主管布鲁斯·弗里德曼博士和核心经理戈登·鲁塞尔博士的专业知识极大地促进了核心的扩大利用。这两家公司都为我们的用户开发了一系列新的应用程序。作为一个团队，我们拥有大量的集体专业知识，包括对大脑、肠道、皮肤和次级淋巴器官中的病原体感染和免疫反应的多光子实时成像，以及组织基质中肿瘤的行为。基于FRET/FLIM的方法被用来跟踪信号分子在受体刺激下的定位、相互作用和活性，光激活以在原位瞬时产生信号中间产物，以及钙和线粒体功能的实时成像。自成立以来，这一核心产生的数据为多份手稿和成功的赠款申请做出了贡献。我们用户组的实验需求不断驱使我们扩展我们的能力，本提案中要求的每个硬件组件都是为了满足我们活跃用户群的特定当前和未来需求而选择的。这项应用的最终目标是使这台仪器现代化，并利用技术发展来增强自安装以来可用的多光子成像能力。所需资金将用于硬件升级，这将1)提高整体能力，包括成像深度、采集速度/时间分辨率、空间覆盖、探测灵敏度和激发灵活性；2)更换控制仪器的传统计算机系统，增加离线流和分析能力；3)为活体组织和细胞的长期实时成像提供强大的环境控制。

 DESCRIPTION (provided by applicant): In response to the expanding need for advanced live cell imaging capabilities, the University of Pennsylvania School of Veterinary Medicine (SOVM) established an Imaging Core facility in the summer of 2008. With the involvement of several pioneer labs, and with nearly $900,000 contributed by the University Provosts office and Departments and Centers in the SOVM, we purchased several microscopes including a Leica SP5 multiphoton system. Expanding utilization of the core has been greatly facilitated by the expertise of our core director, Dr. Bruce Freedman and the core manager, currently Dr. Gordon Ruthel. Both have been instrumental in the development of a range of new applications for our users. As a group we have an enormous amount of collective expertise, including multiphoton real-time imaging of pathogen infection and immune responses in the brain, intestine, skin, and secondary lymphoid organs, as well as the behavior of tumors within the tissue matrix. FRET/FLIM based approaches are used to track the localization, interactions, and activity of signaling molecules in response to receptor stimulation, photo-activation to generate signaling intermediates instantaneously in situ, and live imaging of calcium and mitochondrial function. Since its inception, the data generated in this core has contributed to multiple manuscripts and successful grant applications. The experimental needs of our user group continuously drives us to expand our capability and each of the hardware components requested in this proposal was chosen to address specific current and future needs of our active user base. The ultimate goal of this application is to modernize this instrument and capitalize on technological developments to enhance multiphoton imaging capability that have become available since its installation. Funds requested will be used for hardware upgrades that will 1) increase the overall capability including depth of imaging, acquisition speed/temporal resolution, spatial coverage, detection sensitivity, and excitation flexibility 2) replace a legacy computer system that controls the instrument and add off line streaming and analysis capability, and 3) provide robust environmental control for long-term real time imaging of living tissues and cells.