Two-photon Microscope System for In-vivo Imaging

描述（由申请方提供）：双光子显微镜可用于观察活动物完整器官的三维自然环境中的细胞过程，如细胞迁移、细胞与细胞和细胞与基质的相互作用、肿瘤生长、微生物感染和基因表达。在过去的几年里，双光子显微镜已经彻底改变了我们对发育、稳态和病理过程的看法。我们建议在Wistar研究所建立一个活体双光子显微镜系统，通过收购徕卡TCS SP2光谱共聚焦和双光子显微镜结合光谱物理宽带迈泰飞秒脉冲激光。拟议仪器的主要用户将是一组来自广泛学科的Wistar研究人员，包括免疫学、肿瘤生物学、皮肤生物学和传染病研究。需要将来自培养模型的实验数据转化为活体动物，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Wistar研究所对活体成像的需求。这种研究无法通过现有设备进行。基于有希望的初步研究，研究人员将深入探索免疫系统与肿瘤细胞之间的体内相互作用，炎症和感染性疾病期间的免疫细胞相互作用，以及肿瘤发生和进展期间的黑素细胞/黑色素瘤细胞行为。该仪器将位于研究所的中心位置，方便所有用户使用，将由专门的仪器操作员管理，并由显微镜咨询委员会监督。来自不同领域的研究人员将有新的机会进行互动，并启动跨学科研究项目，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疾病病理机制的新见解，包括自身免疫，癌症和慢性感染。

DESCRIPTION (provided by applicant): Two-photon microscopy allows for the visualization of cellular processes, such as cell migration, cell-to-cell and cell-to-matrix interactions, growth of tumors, microbial infection, and gene 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 Wistar Institute through the acquisition of a Leica TCS SP2 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 Wistar Investigators drawn from a wide range of disciplines, including immunology, tumor biology, skin biology, and infectious disease 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 intravital imaging at the Wistar Institute. Such research cannot be conducted by equipment currently available. Based on promising preliminary studies, the Investigators will explore in-depth the interactions between the immune system and tumor cells in vivo, immune cell interactions during inflammatory and infectious diseases, and melanocyte/melanoma cell behavior during tumor development and progression. The instrument will be centrally located at the Institute to enable convenient access for all users, will be managed by a dedicated instrument operator, and supervised by the 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 pathomechanisms of disease, including autoimmunity, cancer, and chronic infections.