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

用于 2 光子显微镜的 FLIM 系统、共振扫描仪和紫外激光器

• 批准号: 7794472
• 负责人: BRUCE D FREEDMAN
• 金额: $ 49.63万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7794472
• 关键词: Address; Brain; Calcium; Cells; Consultations; Core Facility; Data; Detection; Energy Transfer; Experimental Designs; Fluorescence Resonance Energy Transfer; Funding; Future; Goals; Image; Immune response; Immunity; In Situ; Infection; Intestines; Lasers; Life; Lymphoid; Microscope; Mitochondria; Organ; Paper; Pharmaceutical Preparations; Photons; Preparation; Publications; Resolution; Schools; Signal Transduction; Signaling Molecule; Skin; Staging; System; Time; Tissues; United States National Institutes of Health; Veterinary Medicine; base; cellular imaging; improved; instrument; interest; pathogen; receptor; response

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.

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，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发展了专门和广泛的用户基础，NIH资助的12个实验室积极地从设施中生成数据。此外，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迅速扩大。核心董事张凌利博士的专业知识极大地促进了核心应用的扩大利用，他为我们的用户开发和实施了一系列新的应用程序。作为一个团队，我们已经开发了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实时成像，基于FRET的方法来跟踪信号分子对受体刺激的定位、相互作用和活动，光激活以在原位瞬时生成信号中间体，以及钙和线粒体功能的实时成像。虽然核心系统全面投入使用的时间相对较短，但所产生的数据有助于编写一份豁免文件，其他研究的结果已提交发表或处于最后编写阶段。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特定的当前需求，并预测未来的需求，最终目标是提高我们的双光子/光谱成像能力。因此，所需资金将用于升级硬件，以从3个方面增强我们的核心共焦/双光子光谱成像系统的能力：1)提高显微镜的时间分辨率、空间覆盖范围和检测灵敏度，2)提供药物应用能力和足够的环境控制，以便对活组织和细胞进行长期实时成像，以及3)扩展仪器的整体能力，以执行Forster共振能量转移(FRET)和荧光寿命成像。