4-Laser Cell Sorter

流式细胞仪是一种检测和测量一组细胞或颗粒的物理和化学特征的仪器，提供单个细胞/颗粒水平的可量化数据。建议的仪器包括一个高速细胞分选器，它根据细胞的光学性质对感兴趣的细胞进行物理分离。拟议的流式细胞仪将在多用户概念的框架内辅助分子生物学研究。不同的研究主题包括蓝藻、酵母、植物和哺乳动物细胞的合成生物学，以及对人类细胞的操作和监测，例如β细胞、肝细胞、内皮细胞。糖聚合物生产的合成代谢途径以及糖和激素生物传感器将在单细胞水平上进行工程设计和定量表征，这是合成生物学方法的关键。将建立新的基于细胞仪的方法，如ITRAP方法，该方法开发针对分别与阿尔茨海默病和II型糖尿病相关的多肽的蛋白水解性抗体衍生物。建立锥虫及其变形杆菌内共生菌的可诱导基因敲除体系，并对不同细胞周期阶段进行分离和鉴定。将对高性能光遗传开关进行工程设计，并通过定向进化获得具有定制功能的光遗传开关。此外，利用哺乳动物系统中的多色检测，将建立转基因生物分子的相对量化。基于流式细胞仪的细胞分选将用于根据荧光团的表达来分离例如肠道细胞类型，这将有助于阐明新观察到的肠道干细胞的昼夜节律。总之，拟议的仪器-一个包括四种不同激光/颜色的流式细胞仪和一个高速分选机-将通过提供绝对必要的、用户友好的和非常灵活的基础设施来支持我们部门正在进行的大量研究项目和倡议。

A flow cytometer is an instrument that detects and measures physical and chemical characteristics of a population of cells or particles providing quantifiable data at the single cell/particle level. The proposed instrument includes a high-speed cell sorter, which physically separates cells of interest based on their optical properties. The proposed flow cytometer will assist molecular biology research within the frame of a multi-user concept. Diverse research topics are aimed ranging from synthetic biology in cyanobacteria, yeast, plant and mammalian cells, towards manipulation and monitoring of human cells, e.g. beta cells, hepatocytes, endothelial cells. Synthetic metabolic pathways for sugar polymer production as well as sugar and hormone biosensors will be engineered and quantitatively characterized at the single cell level, which is key in synthetic biology approaches. Novel, cytometer-based methods will be established like the iTRAP method, which develops proteolytic antibody derivatives targeting peptides involved in Alzheimer’s Disease and type II diabetes, respectively. The establishment of an inducible knock out system is aimed for a trypanosomatid and its proteobacterial endosymbiont as well as isolation and characterization of different cell cycle stages. High-performance optogenetic switches will be engineered and optogenetic switches with custom functionality obtained through directed evolution. Further, a relative quantification of transgenetically-implemented biomolecules will be established using multicolor detection in mammalian systems. Flow cytometer based cell sorting will be used to separate e.g. intestinal cell types according to fluorophore expression, which will allow for elucidation of a newly observed circadian rhythm in intestinal stem cells. In summary, the proposed instrument - a flow cytometer including four different lasers/colors and a high speed sorter - will support a plethora of ongoing research projects and initiatives at our department by providing an absolutely essential, user-friendly and very flexible infrastructure.