Multiphoton imaging and uncaging system with a capability of simultaneous electro

描述（由申请人提供）：本申请要求资金用于购买具有成像和释放能力的双光子显微镜系统，该系统将构成圣地亚哥加州大学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，能够以亚微米分辨率对深度达500微米的完整脑组织进行成像。我们已经从UCSD的多个部门召集了一个基础广泛的优秀用户组，主要关注神经系统疾病和癌症动物模型的临床前研究。随着新项目获得资助，以及我们吸引更多的调查人员，我们预计用户群将增长。拟议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，皮层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，最小化光损伤，成像和共同定位多种荧光指示剂的能力，亚细胞光活化和同时电生理记录的能力。所有这些要求都解决了拟议的国家的最先进的双光子显微镜多用户设施。选择所要求的系统是因为其高图像质量、高采集速度、双光子成像和解除锁定光学路径的容易校准以及与电生理组件的兼容性。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，支持笼状化合物的双光子光活化和荧光寿命成像显微镜（FLIM）。 在行政上，拟议的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，并为资源管理、充电机制和用户培训建立了基础设施。拟议的双光子仪器是对中心现有神经成像模式的补充，如MEG，PET和fMRI，并将提供UCSD多模态成像工作的重要组成部分。具体而言，结合功能磁共振成像，PET和体内双光子显微镜将有助于弥合差距之间的非侵入性人类和单细胞动物表征疾病相关的变化。研究人类疾病的动物模型是UCSD医学院正在进行的一项重要工作。 总之，拟议的尖端双光子设施将为UCSD的NIH资助的研究人员提供一个重要的工具，以研究大脑激活、疾病进展特征、疾病诊断和治疗中涉及的复杂途径。这些研究将影响我们对正常和病理性脑功能的理解，为治疗和预防人类疾病开辟新的途径。拟议的设施预计也将在UCSD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.