Multiphoton upgrade of confocal microscope for live organ and tissue imaging

双光子显微镜允许可视化细胞过程，例如细胞 迁移，细胞与细胞和细胞与基质的相互作用，亚细胞器功能 （线粒体），细胞信号（Ca 2+，活性氧和磷脂 摄取），肿瘤生长的表征，药物靶向，和内的蛋白质表达。 活体动物完整器官的三维自然环境。期间 在过去的几年里，双光子显微镜彻底改变了我们对发育的看法， 稳态和病理过程。我们建议建立一个活体双光子 环境医学研究所（IFEM）的显微镜系统， Leica TCS SP 5光谱共聚焦和双光子显微镜的采购 与光谱物理学宽带迈泰飞秒脉冲激光器组合。的 拟议仪器的主要用户将是一组医学院 研究人员来自广泛的学科，包括生理学，病理学， 急诊医学，肺医学，癌症生物学，流变学， 血液学/肿瘤学和免疫学研究。翻译实验的必要性 数据从培养模型转化为活体动物，并将静态信息扩展为动态信息 将细胞过程转化为体内四维动力学分析， IFEM需要活体成像。这样的研究不能由 现有设备。记录ROS的有希望的初步研究 肺中的产生，纳米载体的血管靶向，T细胞祖细胞的运输 在体内，和免疫细胞的相互作用，在炎症和感染性疾病， 证明了双光子显微镜用于拟议项目的可行性。的 仪器将位于医学院的中心，以方便 所有用户的访问，将由IFEM在PI的监督下维护， 显微镜咨询委员会。来自不同领域的研究人员将有新的 互动的机会和跨学科研究项目的启动， 将大大推进他们的NIH资助项目。预计该文书 将有助于获得新的见解正常细胞生理学，药物输送， 涉及衰老、癌症、急性肺损伤、移植 肺动脉高压、动脉粥样硬化、自身免疫和慢性感染。 相关性：双光子显微镜的目的是促进增加相互作用 在临床研究者和基础科学家中， 各种疾病模型及其可能的治疗方法。这个工具将是无价的 将体外细胞培养研究转化为完整器官或活体动物。总体看 设备将增强和扩展IFEM核心，为 宾夕法尼亚大学医学院。

Two-photon microscopy allows for the visualization of cellular processes, such as cell migration, cell-to-cell and cell-to-matrix interactions, subcellular organelle function (mitochondria), cellular signaling (Ca2+, reactive oxygen species, and phospholipid uptake), characterization of tumor growth, drug targeting, and protein expression within the three-dimensional, natural environment of intact organs in living animals. During the past few years, two-photon microscopy has revolutionized our view of developmental, homeostatic, and pathologic processes. We propose to establish an intravital two-photon microscopy system at The Institute for Environmental Medicine (IFEM) through the acquisition of a Leica TCS SP5 Spectral Confocal and Two-Photon Microscope in combination with a Spectra-Physics Broadband Mai-Tai femtosecond-pulsed Laser. The major users of the proposed instrument will be a group of School of Medicine Investigators drawn from a wide range of disciplines, including Physiology, Pathology, Emergency Medicine, Pulmonary Medicine, Cancer Biology, Rheumatology, Hematology/Oncology and Immunology research. The need to translate experimental data from culture models into living animals, and to expand static information of dynamic cellular processes into four- dimensional kinetic analyses in vivo has greatly augmented the need for intra-vital imaging at the IFEM. Such research cannot be conducted by equipment currently available. Promising preliminary studies documenting ROS production in the lung, vascular targeting of nanocarriers, trafficking of T-cell progenitors in vivo, and immune cell interactions during inflammatory and infectious diseases has demonstrated the feasibility of two-photon microscopy for the proposed projects. The instrument will be centrally located in the School of Medicine to enable convenient access for all users, will be maintained by the IFEM under the supervision of the PI and Microscope Advisory Committee. Investigators from diverse fields will have new opportunities for interaction and the initiation of interdisciplinary research projects, which will significantly advance their NIH-funded projects. It is anticipated that the instrument will help to obtain novel insights into the normal cellular physiology, drug delivery, and various pathologies involved with aging, cancer, acute lung injury, transplantation, pulmonary hypertension, atherosclerosis, autoimmunity, and chronic infections. Relevance: The purpose of the 2-Photon microscope is to facilitate increased interaction among clinical investigators and basic scientists who are dedicated towards the study of various disease models and their possible treatment. This instrument will be invaluable in translating in vitro cell culture studies to intact organ or live animal. Overall, the equipment will enhance and expand IFEM Core to provide state of the art support to the School of Medicine at Penn.