Quantitative high-throughput spinning-disk confocal microscope

定量高通量转盘共焦显微镜

• 批准号: 512574446
• 金额: --
• 依托单位: Goethe-Universität Frankfurt am Main
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2023
• 资助国家: 德国
• 起止时间: 2022-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/512574446?language=en
• 关键词: Quantitative; throughput; spinning; disk; confocal

For further equipment of the infrastructure center Frankfurt Center for Innovation & Technologies (FCiT, currently being established) a confocal spinning-disk microscope for high-throughput analyses is requested. Within the FCiT, several life science and biomedical oriented devices in different categories are brought together and operated jointly. In particular, the instruments serve the interface between basic and preclinical research and provide the technological basis for various joint research consortia. These include the Structural Genomics Consortium (EUbOPEN, since 2017), the Innovative Medicine Initiative EUbOPEN (since 2020), the Cluster Initiative ENABLE (since 2021) and the Cluster for Future PROXIDRUGS (since 2021). In contrast to classical cell biology projects, in which individual cellular structures are analyzed by high-resolution confocal microscopy (required technologies provided by the Frankfurt Institute for Advanced Light Microscopy (FCAM)), the above-mentioned collaborative projects require much higher throughput (hardware) and correspondingly optimized and automated analysis capabilities (software). Especially the automated microscopic analysis of cells in microtiter plate formats are crucial here. In the Cellular & Phenotypical Screening Platform (Head: Dr. Alexandra Stolz), chemical libraries are examined microscopically in cell-based, medium- to high-throughput analyses. Analyses start with the examination of single chemical substances in different concentrations and extend to screens of relatively large chemical libraries (e.g. FDA approved drug library of ~3,000 cpds). Advanced, more detailed microscopic analyses is performed subsequently in collaboration with the FCAM. The automated confocal high-content screening microscope with specific software applied for, can scan fixed cells/tissues as well as live cultures in a wide variety of plate formats in quantitative long-term microscopy in a semi-automated manner and process the acquired data by means of intelligent, user-specific logarithms and analysis protocols. Equipment for the screening microscope includes multiple laser and filter combinations, objectives, a high-performance camera, temperature and gas regulation of the incubation chamber, and various software packages for automated image analysis. The scope of the instrument includes various microscopy-based techniques for the analysis of 2D and 3D cultured cells as well as tissues, including those necessary for localization studies, morphology studies, screens for identification, characterization and validation of chemical compounds (drug discovery; PROTACs; small molecules), studies for identification of altered cellular signaling pathways, protein-protein interactions, degradation studies of cellular proteins and cell characterization (cellomics).

对于基础设施中心法兰克福创新与技术中心（FCiT，目前正在建立）的进一步设备，需要用于高通量分析的共聚焦旋转盘显微镜。在FCiT内，几个不同类别的生命科学和生物医学导向的设备被汇集在一起并共同操作。特别是，这些仪器服务于基础研究和临床前研究之间的接口，并为各种联合研究联盟提供技术基础。其中包括结构基因组学联盟（EUbOPEN，自2017年起），创新医学计划EUbOPEN（自2020年起），集群计划ENABLE（自2021年起）和未来proxidmedicines集群（自2021年起）。经典的细胞生物学项目中，单个细胞结构是通过高分辨率共聚焦显微镜分析的（所需技术由法兰克福高级光学显微镜研究所（FCAM）提供），与此相反，上述合作项目需要更高的吞吐量（硬件）和相应的优化和自动化分析能力（软件）。特别是在微滴板格式的细胞的自动显微分析是至关重要的。在细胞和表型筛选平台（负责人：Alexandra Stolz博士）中，化学文库在细胞为基础的中到高通量分析中进行显微镜检查。分析从检查不同浓度的单一化学物质开始，并扩展到相对较大的化学文库的筛选（例如FDA批准的~3,000 cpds的药物文库）。先进的，更详细的微观分析随后与FCAM合作进行。采用专用软件的全自动共聚焦高含量筛选显微镜，可以半自动地扫描固定细胞/组织以及定量长期显微镜中多种平板格式的活培养物，并通过智能、用户特定的对数和分析协议处理获取的数据。筛选显微镜的设备包括多种激光和滤镜组合，物镜，高性能相机，孵育室的温度和气体调节，以及用于自动图像分析的各种软件包。该仪器的范围包括用于分析2D和3D培养细胞以及组织的各种基于显微镜的技术，包括定位研究、形态学研究、鉴定筛选、表征和化合物验证所必需的技术(药物发现；PROTACs；小分子)，研究改变的细胞信号传导途径，蛋白质蛋白质相互作用，细胞蛋白质降解研究和细胞表征（细胞组学）。