Confocal Laser Scanning Microscope (CLSM)

申请人申请共聚焦激光扫描显微镜（CLSM），该显微镜将由微生物学家和感染生物学家自2017年以来在研究大楼“微生物功能基因组学中心”（C_FunGene）使用。C_FunGene主持研究感染生物学和环境微生物学课题的研究小组。申请人的资助项目旨在阐明病原体-宿主相互作用的机制，病原微生物的毒力策略或细菌细胞-细胞通讯。为了避免使用动物感染模型，越来越多地使用细胞培养或基于组织的感染模型来破译病原体与其宿主或特定宿主组织之间的分子和细胞相互作用。这些模型允许实时分析和可视化活体外的感染过程。细菌群落中生物膜形成和细胞间通讯的机制是环境微生物学研究的重点。因为生物膜形成也与人或动物中的细菌定殖和感染相关，所以申请人具有共同的兴趣。CLSM对于实时可视化细胞或类器官系统中的感染过程或生物膜形成至关重要。CLSM将安装在显微镜室中，该显微镜室位于C_FunGene的S2实验室区域。C_FunGene的所有研究小组和其他对感染/免疫调节或细胞生物学感兴趣的小组将根据规定访问CLSM。CLSM将用于回答与感染生物学相关的问题。将在遵守生物安全2级措施的情况下，对临床相关病原体开展工作。重要的是，这台CLSM将是Greifswald S2实验室中唯一一台最先进的共聚焦显微镜。由于CLSM将是一个多用户设备，因此选择了一个非常广泛和现代化的设备，以满足潜水员申请人群体的各种要求。其应用范围从单个细菌细胞和细胞群落的高分辨率显微镜到受感染真核细胞的实时监测。脉冲白色激光器完美匹配任何荧光团。即使在光谱非常接近的情况下，也可以同时使用多达八条激发线，这允许新的实验方法，包括蛋白质定位和共定位研究。对于活细胞成像，所要求的CLSM还配备了一个大型温控孵育室，具有主动气体调节功能。

The applicants apply for a confocal laser scanning microscope (CLSM) that will be used by microbiologists and infection biologists working since 2017 in the research building "Center for Functional Genomics of Microbes" (C_FunGene). C_FunGene hosts research groups working on topics in infection biology and environmental microbiology. The funded projects of the applicants aim to elucidate mechanisms of pathogen-host interactions, virulence strategies of pathogenic microorganisms or bacterial cell-cell communication. To avoid the use of animal infection models, cell culture or tissue-based infection models are increasingly used to decipher molecular and cellular interactions between a pathogen and its host or specific host tissues. These models allow real-time analysis and visualization of the infection process outside of a living organism. The mechanisms of biofilm formation and cell-cell communication in bacterial communities are of high priority in environmental microbiology. Because biofilm formation is also associated with bacterial colonization and infections in humans or animals, the applicants share common interests. The CLSM is pivotal to visualize infection processes in cellular or organoid systems or biofilm formation in real-time. The CLSM will be installed in a microscopy room, which is located in the S2 laboratory area of C_FunGene. All research groups of the C_FunGene and other groups interested in infections/immune modulation or cell biology will have access to the CLSM according to the regulations. The CLSM will be used to answer questions relevant to infection biology. The work will be performed with clinically relevant pathogens under observance of biosafety level 2 measures. Importantly, this CLSM will be the only state-of-the art confocal microscope in Greifswald in an S2 laboratory. Since the CLSM will be a multiple user device, a very broad and modern equipment was selected to be able to meet the various requirements of the divers group of applicants. The applications range from high-resolution microscopy of individual bacterial cells and cell communities to real-time monitoring of infected eukaryotic cells. The pulsed white light laser perfectly matches any fluorophore. Up to eight excitation lines can be used simultaneously, even in case of close proximity of spectra, which allows new experimental approaches including protein localization and co-localization studies. For live cell imaging, the requested CLSM is further equipped with a large, temperature-controlled incubation chamber with active gas regulation.