Two-photon microscope with integrated photostimulation and electrophysiology modules

我们在此申请一台集成光刺激和电生理模块的双光子显微镜，用于德国维尔茨堡大学生理学研究所的神经科学和心血管研究。一个中心应用领域是使用Ca2+敏感指标分析神经网络动力学和突触过程。该显微镜还将研究利钠肽对冠状动脉微循环的影响，以及巨核细胞和血小板的生理和病理生理学。拟议研究项目的指导原则是考虑到生理过程只能在其三维结构的背景下得到充分理解。为此，该系统配备了一个共振-振镜-振镜扫描仪和一个快速压电致动器，以便在高时间和/或空间分辨率下测量生物过程，甚至在多个光学部分。光遗传方法可以通过专用的光刺激模块集成。该系统还允许进行电生理（膜片钳）和光学记录。提出的显微镜具有模块化设计，使光学-电生理应用于活体动物体内和体外制备。所要求的设备也将提供给维尔茨堡大学和维尔茨堡大学医院的其他科学家。由于其独特的技术特点，所提出的双光子显微镜将为活细胞成像开辟新的可能性，从而为加强w<s:1> rzburg的生物医学研究做出重要贡献。

We here apply for a two-photon microscope with integrated photostimulation and electrophysiology modules for neuroscientific and cardiovascular research at the Institute of Physiology of the University of Würzburg. A central field of application is the analysis of neuronal network dynamics and synaptic processes using Ca2+-sensitive indicators. Other scientific topics to be addressed by the proposed microscope include the effects of natriuretic peptides on coronary microcirculation as well as the physiology and pathophysiology of megakaryocytes and platelets. A guiding principle of the proposed research projects is the consideration that physiological processes can only be adequately understood in the context of their three-dimensional structure. To this end, the system is equipped with a resonance-galvanometer-galvanometer scanner and a fast piezo actuator in order to enable measurements of biological processes at high temporal and/or spatial resolution even in multiple optical sections. Optogenetic methods can be integrated via a dedicated photostimulation module. The system also allows combined electrophysiological (patch-clamp) and optical recordings to be performed. The proposed microscope has a modular design, which enables optical-electrophysiological applications in both living animals in vivo and in vitro preparations. The requested device will also be made available to other scientists at the University of Würzburg and the University Hospital of Würzburg. Due to its unique technical features, the proposed two-photon microscope will open up new possibilities for live-cell imaging and thus make an important contribution to strengthening biomedical research in Würzburg.