automated high-content-fluorescence-microscope

点击翻译按钮获取中文摘要

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.