Two-Photon Microscope Patch Clamp Integrated System

描述（由申请人提供）：要求双光子扫描共聚焦显微镜与膜片钳装置集成。这个集成系统将用于结合成像和同时记录活细胞电生理的实验。六名调查员，Ed Chapman, Roberto Coronado, Robert Fettiplace, Meyer Jackson， Jeff Walker和Lea Ziskind-Conhaim构成了该系统主要用户的核心。所有这些研究人员都具有电生理学和成像方面的专业知识，并且都对能够结合激光扫描显微镜和贴片夹的系统有类似的需求。他们将研究心肌细胞、毛细胞、突触前终末和神经内分泌终末的信号传导的各个方面。钙在这些系统的生理机能中起着至关重要的作用。大多数实验主要通过钙成像进行，以获得细胞受到电刺激时钙信号的实时高分辨率信息。这六名研究人员中的每一位都有美国国立卫生研究院的研究资金，这些研究可以通过该仪器得到极大的提高。除此之外，该系统将使Chapman能够研究synaptotagmin对融合孔的调节，Coronado可以研究单个ca2 +释放位点的ca2 +动力学，Fettiplace可以研究单个立体纤毛中的ca2 +动力学，Jackson可以研究神经末端ca2 +动力学的空间变化，Walker可以将笼状信号分子应用于t小管，ziskin - conhaim可以研究特定细胞类型在脊髓节律性中的作用。共享仪器拨款将通过在ca2 +成像中提供更好的信噪比，在切片成像实验中更好的组织穿透，以及在笼状化合物光解中更好的定位，来加强所有这些研究项目。

DESCRIPTION (provided by applicant): A two-photon scanning confocal microscope is requested for integration with a patch clamp setup. This integrated system will be used for experiments that combine imaging with simultaneous electrophysiological recording from living cells. Six investigators, Ed Chapman, Roberto Coronado, Robert Fettiplace, Meyer Jackson, Jeff Walker, and Lea Ziskind-Conhaim form a core of major users of this system. All of these investigators have expertise in both electrophysiology and imaging, and all have similar needs for a system that can combine laser scanning microscopy and patch clamping. They will investigate various aspects of signaling in cardiac myocytes, hair cells, presynaptic terminals, and neuroendocrine terminals. Calcium plays vital roles in the physiology of each of these systems. Most of the experiments to be conducted primarily with calcium imaging to obtain high resolution information about calcium signals in real time as cells are subject to electrical stimulation. Each of the six investigators has NIH funding for research that can be dramatically enhanced by this instrument. Among other advantages, this system will enable Chapman to investigate the regulation of fusion pores by synaptotagmin, Coronado to investigate Ca 2+dynamics at individual Ca 2+release sites, Fettiplace to study Ca 2+ in individual stereocilia, Jackson to investigate spatial variations in Ca 2+dynamics in nerve terminals, Walker to apply caged signaling molecules to T-tubules and Ziskind-Conhaim to investigate the role of specific cell types in spinal rhythmicity. The shared instrument grant will enhance all of these research programs by providing better signal-to-noise in Ca 2+ imaging, better tissue penetration in imaging experiments in slices, and better localization in caged compound photolysis.