Multiphoton Confocal Microscopy System

描述（由申请人提供）：了解神经系统的发育和功能需要阐明神经回路的独立组成部分及其在完整系统中的功能之间的关系。在完整的系统中揭示这些关系已被证明是非常困难的传统方法。然而，多光子共聚焦显微镜的出现，为神经组织深处的体内光学成像提供了最小光损伤的机会。该方法可用于特定细胞类型的靶电极记录或光学探测已识别细胞类型的活性。神经元活动的光学监测可以通过测量钙敏感染料的荧光变化或神经递质突触前囊泡释放的基因表达传感器来完成。在本提案中，我们计划使用多光子共聚焦显微镜，结合遗传编码光学报告和其他小荧光分子，研究正常成人神经系统中神经活动的连通性和作用，以及在发育和疾病状态下突触的形成。这些研究只有使用多光子共聚焦显微镜系统才有可能

DESCRIPTION (provided by applicant): Understanding nervous system development and function requires elucidation of the relationships between the separate components of neural circuits and their function in an intact system. Revealing these relationships in intact systems has proven to be extremely difficult with conventional methods. The advent of multiphoton confocal microscopy, however, provides the opportunity for in vivo optical imaging deep within neuronal tissue with minimal photodamage. This method can be used to target electrode recordings to specific cell types or to optically probe the activity of identified cell types. Optical monitoring of neuronal activity can be done using measurements of fluorescent changes from calcium sensitive dyes or genetically expressed sensors of presynaptic vesicular release of neurotransmitter. In the present proposal, we plan to use multiphoton confocal microscopy, coupled with genetically encoded optical reporters and other small fluorescence molecules, to study the connectivity and the role of neural activity in the normal adult nervous system and in the formation of synapses during development and in disease states. These studies are only possible with the use of a multiphoton confocal microscopy system