Multiphoton upgrade of confocal microscope for live organ and tissue imaging

用于活体器官和组织成像的共焦显微镜的多光子升级

• 批准号: 7212206
• 负责人: MADESH MUNISWAMY
• 金额: $ 50万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-22 至 2008-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7212206
• 关键词: Acute Lung Injury; Advisory Committees; Aging; Animals; Arts; Atherosclerosis; Autoimmunity; Blood Vessels; Cancer Biology; Cell Communication; Cell physiology; Cells; Chronic; Clinical Investigator; Communicable Diseases; Cultured Cells; Data; Development; Discipline; Disease model; Drug Delivery Systems; Emergency Medicine; Environment; Environmental Medicine; Equipment; Four-dimensional; Funding; Hematology; Image; Imagery; Immune; Immunology; In Vitro; Infection; Inflammatory; Institutes; Interdisciplinary Study; Kinetics; Lasers; Life; Lung; Malignant Neoplasms; Microscope; Microscopy; Mitochondria; Modeling; Organ; Organelles; Pathologic Processes; Pathology; Phospholipids; Photons; Physics; Physiologic pulse; Physiology; Production; Pulmonary Hypertension; Pulmonology; Pulse taking; Purpose; Range; Reactive Oxygen Species; Research; Research Personnel; Research Project Grants; Rheumatology; Scientist; Signal Transduction; Supervision; System; T-Lymphocyte; Tissues; Translating; Transplantation; cell motility; in vivo; insight; instrument; medical schools; nanocarrier; novel; oncology; progenitor; protein expression; trafficking; tumor growth; two-photon; uptake

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.

双光子显微镜可以实现细胞过程的可视化，例如细胞 迁移、细胞间和细胞间相互作用、亚细胞器功能 （线粒体）、细胞信号传导（Ca2+、活性氧和磷脂） 摄取）、肿瘤生长特征、药物靶向和蛋白质表达 活体动物完整器官的三维自然环境。期间 过去几年，双光子显微镜彻底改变了我们对发育、 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 购买 Leica TCS SP5 光谱共焦和双光子显微镜 与 Spectra-Physics 宽带 Mai-Tai 飞秒脉冲激光器相结合。这 拟议仪器的主要用户将是一群医学院 研究人员来自各个学科，包括生理学、病理学、 急诊医学、肺科、癌症生物学、风湿病学、 血液学/肿瘤学和免疫学研究。需要翻译实验 将培养模型的数据转化为活体动物，并扩展动态的静态信息 将细胞过程转化为体内四维动力学分析大大增强了 IFEM 进行活体成像的需求。此类研究不能由 目前可用的设备。记录 ROS 的有希望的初步研究 肺部生产、纳米载体的血管靶向、T 细胞祖细胞的运输 体内和炎症和传染病期间免疫细胞的相互作用 证明了双光子显微镜对于拟议项目的可行性。这 仪器将位于医学院的中心位置，以方便 所有用户的访问权限将由 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获得新的 互动和启动跨学科研究项目的机会， 将显着推进其 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、药物输送和 与衰老、癌症、急性肺损伤、移植有关的各种病理， 肺动脉高压、动脉粥样硬化、自身免疫和慢性感染。 相关性：2 光子显微镜的目的是促进增强互动 致力于研究的临床研究人员和基础科学家 各种疾病模型及其可能的治疗方法。这台仪器将是无价的 将体外细胞培养研究转化为完整器官或活体动物。总体而言， 设备将增强和扩展 IFEM 核心，为 宾夕法尼亚大学医学院。