Super resolution microscope for live cell microscopy

我们用细胞生物学和生物化学的方法研究了bcl2蛋白家族对细胞凋亡和生存决策的动态调控过程，其中蛋白质构象的变化和线粒体膜外膜多蛋白复合体的形成是至关重要的。我们通过分子内链的稳定来研究瞬时蛋白质构象。活细胞的高分辨率显微镜研究对于分析bcl2家族多蛋白复合体的形成、参与信号过程和调控是必不可少的。这项研究是基于莱比锡大学生命科学在战略性跨学科研究概况领域“治疗和诊断中的分子和细胞通信”和“文明疾病”方面的密集合作。对于理解动态平衡和重要的病理学，如肿瘤疾病，具有突出意义的知识的获得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膜外关键结构的显微镜，以便在国际上可见的水平上进行研究。分析活细胞中涉及的细胞结构和蛋白质复合体的动力学、排列和亚结构需要超分辨率显微镜。由于分辨率低和信噪比不足，传统的显微方法不能完成这项任务。超分辨率显微镜的意向收购将决定性地推动莱比锡课题组的研究联网。因此，我们要求集成显微镜系统，以满足显微镜研究的高标准，并通过标准化的工作流程为所有实验要求提供高效、轻松的培训、操作和维护。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。为了我们团队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的S2实验室中。我们还将确保仪器的运行达到最高标准，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率和数据管理。通过融入BCF，我们一方面将受益于出色的平台和技术支持，另一方面将通过研讨会和研讨会分享我们的专业知识。从长远来看，计划将该文书纳入一项合作提案，目标是在VMF建立一所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.