Confocal microscope

运动的控制、对环境信息的反应、记忆…的形成我们所有的活动都依赖于神经系统的功能。神经系统在进化过程中获得的非凡能力，以及支持这些活动的能力，依赖于神经系统中单个细胞的精确架构及其相互作用来形成功能电路。个体神经元的结构和神经元之间的连接在个体发育过程中出现，它们可以在动物成年后经历可塑性变化，在老年动物中可以被破坏或拆解。我们研究神经元如何在发育过程中阐述其复杂的和神经元类型的特定形态。这些都是动态的过程，具有复杂的空间和时间维度，只有在体内才能解决。为了阐明它们的细胞和分子机制，我们利用果蝇黑腹果蝇的发育时间短、幼虫角质层透明以及易于遗传操作的优势。为了可视化细胞动力学方面，我们为我在RWTH新设立的教授职位申请了一台共聚焦显微镜。所要求的显微镜将支持我们的工作，记录发生在体内的神经元在几秒钟内的形态变化，以及在这些过程中分子玩家的动态亚细胞分布。为了能够对细胞动力学进行长期的活体成像，该设置必须结合高空间和时间分辨率，以及高灵敏度的检测，以最大限度地减少动物所受的光照。除了对发育中的动物的单个细胞进行分析外，我们还研究了神经元如何相互作用形成功能电路。这些相互作用的三维结构还需要通过压电控制的z步进进行快速体积扫描。为了研究回路中神经元之间的功能联系，以及在变性条件下神经元的可塑性和电位退化，我们进一步阐明了回路中突触的分子组织。考虑到突触的大小和其中单个分子成分的相对距离，所要求的设置将提供同时具有多个探测通道的超分辨率成像。这台显微镜将成为我在RWTH实验室的几乎每一个项目的关键仪器。因此，它对提案中描述的广泛应用的适应性，以及它的易维护性和可获得的支持，将是至关重要的。

The control of movement, the response to environmental information, the formation of memories… all of our activities depend on the function of the nervous system. The extraordinary capacities that the nervous system has acquired during evolution and that support these actions rely on the precise architecture of the individual cells in the nervous system and of their interactions to form functional circuits. The structure of individual neurons and the connections among neurons into ensembles emerge during the individual’s development, they can undergo plastic changes into the adult life of the animal and can be damaged or disassembled in the aged animal. We study how neurons elaborate their complex and neuron-type specific morphologies during development. These are dynamic processes with complex spatial and temporal dimensions that can only be tackled in vivo. To elucidate their cellular and molecular mechanisms, we thus utilize the fruit fly Drosophila melanogaster, taking advantage of its short developmental time, the transparency of the larval cuticle- and the easy of genetic manipulation. To visualize the cellular dynamics aspects, we request a confocal microscope for my newly established Professorship at the RWTH. The requested microscope will support our work recording the morphological changes of neurons happening in vivo within seconds, as well as the dynamic subcellular distribution of molecular players during these processes. To allow long-term in vivo imaging of cellular dynamics, it is essential that the setup combines high spatial and temporal resolution, with highly sensitive detection to minimize the illumination sustained by the animal. In addition to the analysis of individual cells in the developing animal, we investigate how neurons interact with each other to form functional circuits. The three-dimensional architecture of these interactions requires in addition rapid volumetric scans allowed by a piezo-controlled z stepping. To investigate the functional connections among neurons in circuits, their plasticity and potential degradation in degenerative conditions, we furthermore elucidate the molecular organization of synapses within circuits. Given the size of synapses and the relative distance of individual molecular components within them, the setup requested will afford superresolution imaging with multiple detection channels simultaneously. This microscope will be a key instrument for almost every project in my lab at the RWTH. Therefore, its adaptability to the broad spectrum of applications described in the proposal, as well as its easy maintenance and the presence of reachable support will be of vital importance.