Multiphoton Confocal Microscopy for Biomedical Research

描述（由申请人提供）：多光子显微镜是一种强大的新工具，有助于在活体动物完整器官的三维自然环境中研究离散细胞过程。在过去的几年里，多光子显微镜已经彻底改变了我们对发育、体内平衡和病理过程的看法。我们建议通过收购卡尔蔡司LSM 710共聚焦双光子显微镜，在天普大学医学院建立一个活体多光子显微镜系统，该服务目前在校园内不可用。该仪器的主要用户将是一组来自广泛学科的医学院研究人员，包括生物化学、生理学、病理学、药理学、解剖学和细胞生物学、肺部医学、癌症研究所和风湿病学。需要将实验数据从培养模型转化为活体动物，并将动态细胞过程的静态信息扩展为体内四维动力学分析，这极大地增加了坦普尔对生命内成像的需求。这种研究不能用现有的设备进行。有希望的初步研究记录了肺中ROS的产生，内皮祖细胞的血管靶向，体内脑血管通透性以及炎症和感染性疾病期间免疫细胞的相互作用，这些都证明了多光子显微镜在拟议项目中的可行性。该仪器将位于医学院的中心，以便所有用户都能方便地使用；将由生物化学系在PI和显微镜咨询委员会的监督下进行维护。来自不同领域的研究人员将有新的机会进行互动和发起跨学科的研究项目，这将大大推进他们的nih资助的项目。预计该仪器将有助于获得对正常细胞生理学、祖细胞归巢以及与衰老、癌症、急性器官损伤、骨重塑、移植、肺动脉高压、动脉粥样硬化、自身免疫和慢性感染有关的各种病理的新见解。

DESCRIPTION (provided by applicant): Multiphoton microscopy is a powerful new tool that facilitates the study of discrete cellular processes within the three-dimensional, natural environment of intact organs in living animals. During the past few years, multi- photon microscopy has revolutionized our view of developmental, homeostatic, and pathologic processes. We propose to establish an intravital multiphoton microscopy system at Temple University School of Medicine through the acquisition of a Carl Zeiss LSM 710 Confocal and Two-Photon Microscope, a service which is currently unavailable on campus. The major users of the proposed instrument will be a group of School of Medicine Investigators drawn from a wide range of disciplines, including Biochemistry, Physiology, Pathology, Pharmacology, Anatomy and Cell Biology, Pulmonary Medicine, Cancer Research Institute and Rheumatology. 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 Temple. Such research cannot be conducted by equipment currently available. Promising preliminary studies documenting ROS production in the lung, vascular targeting of endothelial progenitors, brain vascular permeability in vivo, and immune cell interactions during inflammatory and infectious diseases has demonstrated the feasibility of multi-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 Biochemistry Department 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, homing of progenitors, and various pathologies involved with aging, cancer, acute organ injury, bone remodeling, transplantation, pulmonary hypertension, atherosclerosis, autoimmunity, and chronic infections.