automated high-content-fluorescence-microscope

我们申请了一台自动化的高通量显微镜，用于细胞培养实验的荧光系统和定量分析。该显微镜将成为<s:1>茨堡大学生物中心核心单元显微镜的一部分。在过去，高通量分析，如siRNA文库的系统分析，是生物化学和分子生物学部门工作的核心部分，是由DFG或欧盟资助的几个合作联盟的一部分。在大多数实验中，使用靶向siRNA文库，例如针对所有泛素连接酶或与MYC癌蛋白相关的所有蛋白质的文库，并研究其对细胞参数的影响。在第二方面的研究中，我们建立了一个系统的细胞应激试验管道（如细胞周期阻滞、细胞凋亡、衰老、内质网应激），这样就可以系统地评估特定蛋白质消耗或药物治疗的效果。该仪器和核心单元被来自生物中心、大学和大学诊所的许多小组的研究人员使用。在过去的几年里，该部门已经建立了与肿瘤类器官有关的工作。特别是，对结肠肿瘤类器官的分析，无论是来自遗传定义的小鼠模型还是来自原发性患者活检，现在已经成为一种常规方法。作为新米尔德里德·谢尔癌症研究中心的一部分，工作还将集中在头颈部鳞状癌类器官与免疫系统细胞的相互作用分析上。所需的三维成像技术需要新的远距离目标和增加的计算能力。第二个新重点将是化合物文库的分析，例如，旨在发现干扰MYC和MYCN癌蛋白转录控制的新物质或破坏转录与DNA复制协调的化合物。为了实现这一目标，我们还优化了邻近结扎法及其定量分析的使用；因此，现在有可能系统地捕捉实验扰动的影响，例如蛋白质的消耗，对细胞内蛋白质复合物的组装和组成。

We apply for an automated high-throughput microscope for the fluorescence-based systematic and quantitative analysis of cell culture experiments. The microscope will be part of the core unit microscopy of the Biocentre of the University Würzburg.In the past, high-throughput analyses such as the systematic analysis of siRNA libraries was central part of the work in the department of biochemistry and molecular biology and was part of several co-operative consortia funded by the DFG or the European Union. In most experiments, targeted siRNA libraries were used such as libraries targeting all ubiquitin ligases or all proteins associated with the MYC oncoprotein and their effects on cellular parameters were investigated. In a second line of research, we built up a systematic pipeline of cellular stress assays (e.g. cell cycle arrest, apoptosis, senescence, ER stress) such that the effect of depletion of a specific protein or a pharmacological treatment could be systematically evaluated. The instrument and the core unit were used by researchers from many groups of the Biocentre, the university and the university clinic.In the last years, work with tumor organoids has been established at the department. In particular, the analysis of colon tumor organoids that are either derived from genetically defined mouse models or from primary patient biopsies is now established as a routine. As part of the new Mildred Scheel Cancer Research Center, work will also focus on the analysis of the interaction of organoids from head-and-neck squamous carcinomas with cells of the immune system. The required three-dimensional imaging techniques necessitate new long-distance objectives and increased computing capacities. A second new focus will be the analysis of compound libraries, for example with the aim to find new substances that interfere with the transcriptional control by the MYC and MYCN oncoproteins or compounds that disrupt the co-ordination of transcription with DNA replication. To achieve this, we have also optimized the use of proximity-ligation assays and their quantitative analysis; as a result, it is now possible to systematically capture the effect of experimental perturbations, such as a depletion of a protein, on the assembly and composition of intracellular protein complexes.