Multiphoton imaging and uncaging system with a capability of simultaneous electro

描述（由申请人提供）：本申请申请资金用于购买具有成像和释放功能的双光子显微镜系统，该系统将构成加州大学圣地亚哥分校神经科学和放射学系的共享设施。双光子显微镜正在成为生物医学研究的重要工具，它提供了对完整脑组织成像的能力，深度可达500微米，分辨率为亚微米。我们汇集了来自加州大学圣地亚哥分校多个部门的优秀的基础广泛的用户群体，主要关注神经疾病和癌症动物模型的临床前研究。随着新项目得到资助，随着我们吸引更多的研究人员，我们希望用户基础能够增长。拟开展的研究涉及神经血管耦合、中风、帕金森病和阿尔茨海默病、神经再生、损伤修复、皮质通讯和发育神经科学。这些研究项目需要光在组织中的深度穿透，光损伤的最小化，成像和共同定位多个荧光指标的能力，亚细胞光激活和同时电生理记录的能力。所有这些要求都是由提议的最先进的双光子显微镜多用户设施解决的。该系统具有成像质量高、采集速度快、双光子成像容易标定、光通道畅通、与电生理元件兼容等特点。该系统允许在体内、体外和离体条件下对结构和功能进行多功能成像，支持笼化化合物的双光子光激活和荧光寿命成像显微镜（FLIM）。行政上，该仪器将成为加州大学圣地亚哥分校功能磁共振成像中心的一部分。该中心为大圣地亚哥地区的用户提供全面的成像设施，并建立了资源管理、充值机制和用户培训的基础设施。拟议的双光子仪器是对中心现有的神经成像模式（如MEG， PET和fMRI）的补充，并将为加州大学圣地亚哥分校多模态成像工作提供重要和核心的组成部分。具体来说，结合fMRI， PET和体内双光子显微镜将有助于弥合非侵入性人类和单细胞动物疾病相关变化表征之间的差距。研究人类疾病的动物模型是加州大学圣地亚哥分校医学院正在进行的一项主要工作。总之，拟议中的尖端双光子设施将为加州大学圣地亚哥分校的美国国立卫生研究院资助的研究人员提供一个重要工具，用于研究涉及大脑激活的复杂途径、疾病进展的特征、疾病诊断和治疗。这些研究将影响我们对正常和病理脑功能的理解，为治疗和预防人类疾病开辟新的途径。拟议中的设施也有望在加州大学圣地亚哥分校的培训和教学工作中发挥重要作用。

DESCRIPTION (provided by applicant): This application requests funds for purchase of a two-photon microscopy system with imaging and uncaging capabilities that will constitute a shared facility in the Departments of Neurosciences and Radiology at the University of California, San Diego. Two-photon microscopy is becoming an important tool for biomedical research, providing the ability to image intact brain tissue to depths of up to >500 microns, with sub-micron resolution. We have assembled an outstanding broadly based group of Users from multiple departments at UCSD with a major focus on pre-clinical studies in animal models of neurological conditions and cancer. We expect the User base to grow, as new projects get funded, and as we attract additional investigators. The proposed research involves neurovascular coupling, stroke, Parkinson's and Alzheimer's diseases, nerve regeneration, injury repair, cortical communication and developmental neuroscience. These research projects require deep penetration of light in tissue, minimization of photodamage, ability to image and co-localize multiple fluorescent indicators, sub-cellular photoactivation and capability of simultaneous electrophysiological recordings. All these requirements are addressed by the proposed state-of-the-art two-photon microscopy multi-user facility. The requested system was chosen because of its high image quality, high acquisition speed, easy calibration of the two-photon imaging and uncaging optical pathways and compatibility with electrophysiology components. The system allows versatile imaging of the structure and function under in vivo, in vitro and ex vivo conditions, supports two-photon photoactivation of caged compounds and Fluorescent Lifetime Imaging Microscopy (FLIM). Administratively, the proposed instrument will be part of The Center for fMRI at UCSD. The Center provides comprehensive imaging facilities for users in the greater San Diego, with established infrastructure for resource administration, recharge mechanisms, and user training. The proposed two-photon instrument is complementary to the existing neuroimaging modalities at the Center such as MEG, PET and fMRI, and will provide an essential and central component of the UCSD Multimodality Imaging effort. Specifically, combining fMRI, PET and in vivo two-photon microscopy will facilitate bridging the gap between non-invasive human and single-cell animal characterization of disease-related changes. Studying of animal models of human diseases is a major ongoing effort at UCSD School of Medicine. In summary, the proposed cutting-edge two-photon facility will provide NIH-funded investigators at UCSD with an essential tool to study the intricate pathways involved in brain activation, features of disease progression, disease diagnosis and treatment. These studies will impact our understanding of normal and pathological brain function, opening new avenues for treatment and prevention of human disease. The proposed facility is also expected to play a significant role in the training and teaching efforts at UCSD.