Multiphoton Confocal Microscopy System

描述(由申请人提供)：要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与其在完整系统中的功能之间的关系。事实证明，用传统方法很难在完整的系统中揭示这些关系。然而，多光子共聚焦显微镜的出现为体内光学成像提供了机会，使其能够以最小的光损伤深入神经元组织。这种方法可用于将电极记录定向到特定的细胞类型或光学探测已识别的细胞类型的活动。通过测量钙敏感染料或突触前囊泡释放神经递质的基因表达传感器的荧光变化，可以完成对神经元活动的光学监测。在目前的计划中，我们计划使用多光子共聚焦显微镜，结合遗传编码的光学记者和其他小荧光分子，来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。这些研究只有在使用多光子共聚焦显微镜系统的情况下才有可能

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