Super resolution microscope for live cell microscopy

用于活细胞显微镜检查的超分辨率显微镜

• 批准号: 514686987
• 金额: --
• 依托单位: Universität Leipzig
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2022
• 资助国家: 德国
• 起止时间: 2021-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/514686987?language=en
• 关键词: Super; resolution; microscope; live; cell

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.

我们调查的动态调节过程中的细胞凋亡和生存的BCL-2蛋白家族使用细胞生物学和生物化学方法，其中蛋白质构象的变化和形成的多蛋白复合物在线粒体外膜是至关重要的。我们研究瞬时蛋白质构象，通过稳定与分子内拴系。活细胞的高分辨率显微镜研究对于分析BCL-2家族多蛋白复合物的形成、参与信号传导过程和调节至关重要。这项研究是基于莱比锡大学生命科学在战略跨学科研究领域“治疗和诊断中的分子和细胞通讯”和“文明疾病”的密切合作。对于理解稳态和重要病理学（如肿瘤疾病）具有突出意义的知识增益越来越需要活细胞的先进成像的支持。因此，迫切需要一种能够解析线粒体外膜关键结构的显微镜，以进行国际可见水平的研究。对活细胞中所涉及的细胞结构和蛋白质复合物的动力学、排列和亚结构的分析需要超分辨率显微镜。传统的显微镜方法不能有助于这项任务，由于缺乏分辨率和信噪比不足。对超分辨率显微镜的收购将决定性地促进莱比锡研究小组的研究网络。因此，我们要求一种集成的显微镜系统，能够满足显微镜研究的高标准，并通过标准化的工作流程实现高效且简单的培训、操作和维护，以满足所有实验要求。由于活细胞成像时间长（数小时），显微镜主要由AG Edlich使用。对于我们小组的前瞻性研究和兽医学院内病原体-宿主关系的密集合作项目，显微镜将位于安全级别S2实验室。我们还将通过将其纳入卓越的生物成像核心设施（BCF），确保仪器以最高标准运行，从而确保仪器利用率和数据管理。通过融入BCF，我们一方面将受益于卓越的平台和技术支持，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，计划将该仪器纳入合作提案，目标是在VMF建立DFG研究生院。