Super resolution microscope for live cell microscopy

我们使用细胞生物学和生化方法研究了BCL-2蛋白家族在细胞凋亡和存活之间的动态调控过程，其中蛋白质构象的变化和线粒体外膜多蛋白复合物的形成是至关重要的。我们通过分子内链的稳定来研究蛋白质的瞬态构象。活细胞的高分辨率显微研究对于分析BCL-2家族多蛋白复合物的形成、参与信号过程和调控至关重要。这项研究基于莱比锡大学生命科学部门在“治疗和诊断中的分子和细胞通讯”和“文明疾病”战略跨学科研究领域的密切合作。知识的增长对于理解体内平衡和重要的病理（如肿瘤疾病）有着重要的意义，越来越需要先进的活细胞成像的支持。因此，迫切需要一种能够分辨线粒体外膜关键结构的显微镜进行国际可见水平的研究。活细胞中涉及的细胞结构和蛋白质复合物的动力学、排列和亚结构分析需要超分辨率显微镜。由于缺乏分辨率和信噪比不足，传统的显微镜方法无法完成这项任务。超分辨率显微镜的预定收购将决定性地促进莱比锡研究小组的研究网络。因此，我们需要一个集成的显微镜系统，以满足高标准的显微镜研究，并允许有效和容易的培训，操作和维护，通过标准化的工作流程，为所有的实验要求。由于活细胞成像时间长（几个小时），该显微镜主要由AG Edlich使用。为了我们小组的前瞻性研究和兽医学院内关于病原体-宿主关系的密集合作项目，显微镜将放置在安全级别为S2的实验室。我们还将确保仪器的操作达到最高标准，将其纳入优秀的生物成像核心设施（BCF），从而确保仪器的利用和数据管理。通过加入BCF，我们一方面将受益于优秀的平台和技术支持，另一方面将通过研讨会和讲习班分享我们的专业知识。从长远来看，计划将该仪器纳入合作提案，目标是在VMF建立DFG研究生院。

We investigate the dynamic regulatory processes of the cellular decision between apoptosis and survival by the BCL-2 protein family using cell biological and biochemical methods, where changes in protein conformations and formation of multiprotein complexes at the outer mitochondrial membrane are of paramount importance. We study transient protein conformations through stabilization with intramolecular tethers. High-resolution microscopic studies of living cells are essential to analyze formation, involvement in signaling processes, and regulation of BCL-2 family multiprotein complexes. This research is based on the intensive collaboration of the life sciences at the University of Leipzig in the strategic interdisciplinary research profile areas "Molecular and Cellular Communication in Therapy and Diagnostics" and "Civilization Diseases". Knowledge gains with outstanding implications for the understanding of homeostasis and significant pathologies, such as tumor diseases, increasingly require the support of advanced imaging of living cells. Therefore, a microscope with the ability to resolve the crucial structures at the outer mitochondrial membrane is urgently needed for research at an internationally visible level. Analysis of the dynamics, arrangement, and substructure of the involved cellular structures and protein complexes in living cells requires super-resolution microscopy. Conventional microscopy methods can not contribute to this task due to lack of resolution and insufficient signal-to-noise ratio. The intended acquisition of the super-resolution microscope will decisively promote the research networking of the research group in Leipzig. We therefore request an integrated microscopy system that meets the high standards of microscopic investigations and allows efficient and easy training, operation, and maintenance through a standardized workflow for all experimental requirements. Due to long live cell imaging times (several hours), the microscope is mainly used by AG Edlich. For perspective research of our group and intensive collaboration projects on pathogen-host relationship within the Faculty of Veterinary Medicine, the microscope will be located in a safety level S2 laboratory. We will also ensure operation of the instrument to the highest standards by incorporating it into the excellent Bioimaging Core Facility (BCF), thereby ensuring instrument utilization and data management. By integrating into the BCF, we will on the one hand benefit from the excellent platform and technical support and on the other hand share our expertise through seminars and workshops. In perspective, it is planned to integrate the instrument into a collaborative proposal with the goal to establish a DFG graduate school at the VMF.