FLIM system, resonant scanner, and UV laser for 2 photon microscope

描述（由申请人提供）：为了满足对先进活细胞成像能力不断扩大的需求，兽医学院建立了一个成像核心设施，该设施于2008年夏天上线。在几个先锋实验室的大力参与下，我们已经实施了一系列尖端应用程序，并在不到一年的时间里，与12个NIH资助的实验室一起，开发了一个专门的、广泛的用户群，积极从该设施生成数据。此外，有7个实验室正处于咨询和实验设计阶段，表明对利用核心设施的兴趣正在迅速扩大。我们的核心董事张玲丽博士的专业知识极大地促进了核心的扩大利用，他在为我们的用户开发和实施一系列新应用程序方面发挥了重要作用。作为一个团队，我们已经开发了大量的集体专业知识，包括大脑、肠道、皮肤和次级淋巴器官中病原体感染和免疫反应的实时成像，基于FRET的方法来跟踪受体刺激下信号分子的定位、相互作用和活性，光激活以即时原位产生信号中间体，以及钙和线粒体功能的实时成像。虽然核心系统全面运作的时间相对较短，但所产生的数据已用于编写一份《豁免》文件，其他研究的结果已提交发表或正处于最后准备阶段。我们用户组的实验需求不断推动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特定的当前需求而选择的，同时也预测了未来的需求，最终目标是提高我们的2光子/光谱成像能力。因此，所要求的资金将用于升级硬件，这将从三个方面增强我们的核心共聚焦/2光子光谱成像系统的能力；1)提高了显微镜的时间分辨率、空间覆盖范围和检测灵敏度；2)为活体组织和细胞的长期实时成像提供了药物应用和足够的环境控制能力；3)扩大了仪器的整体能力，可以进行福斯特共振能量转移（FRET）和荧光寿命成像。

DESCRIPTION (provided by applicant): In response to the expanding need for advanced live cell imaging capabilities, the School of Veterinary Medicine established an Imaging Core facility, which came online in the summer of 2008. With the heavy involvement of several pioneer labs, we have implemented a range of cutting edge applications and in less than one year have developed a dedicated and broad user base with 12 NIH funded labs actively generating data from the facility. In addition, 7 labs are in the consultation and experimental design stage, indicating a rapidly expanding interest in utilizing the core facility. Expanding utilization of the core has been greatly facilitated by the expertise of our core director, Dr. Lingli Zhang, who has been instrumental in developing and implementing a range of new applications for our user. As a group we have developed an enormous amount of collective expertise, including real-time imaging of pathogen infection and immune responses in the brain, intestine, skin, and secondary lymphoid organs, FRET based approaches 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. Although the core has been fully operational for a relatively short time, data generated have contributed to an Immunity paper, and results of other studies have been submitted for publication or are in final stages of preparation. 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 needs, and also to anticipate future needs, with the ultimate goal of improving our 2 photon/spectral imaging capability. Funds requested will therefore be used for upgraded hardware that will enhance the capability of our core confocal/2 photon spectral imaging system in 3 general ways; 1) increase the temporal resolution, spatial coverage, and detection sensitivity of the microscope, 2) provide capability for drug application and adequate environmental control for long term real time imaging of living tissues and cells, and 3) expand the overall capability of the instrument to perform Forster resonance energy transfer (FRET) and fluorescence lifetime imaging.