Flow cytometer with up to five lasers, spectral analyses and auto-sampling

我们的研究试图了解质膜信号通路的空间组织和组装动力学如何控制T细胞的激活和功能。这项研究是基于我们的发现，即质膜蛋白定位于纳米尺寸的区域。我们的研究表明，T细胞的激活是由不同纳米结构域的空间关系、信号复合体在其中的组装以及信号分子在它们之间的移位动态控制的。我们使用多学科策略研究潜在的分子机制，这些策略将单分子显微镜与生化、生物物理和细胞生物学方法相结合。特别是，我们感兴趣的是T细胞受体(TCR)和抑制程序细胞死亡蛋白(PD-1)在T细胞激活和抗肿瘤反应中的途径。我们的最终目标是识别T细胞信号通路中的新控制原理，并利用它们来改变T细胞对未来免疫治疗的反应。我们需要一台带有光谱分析仪和自动进样器的流式细胞仪，这对我们在阿尔伯特-路德维希-弗赖堡大学新建立的实验室以及实现我们当前和未来的研究目标至关重要。所需的流式细胞仪具有多种用途，包括：(I)研究免疫反应和肿瘤环境中免疫细胞的多样性。(Ii)分析T细胞的发育、分化和活动状态。(Iii)原代和转基因细胞的质量控制，以最大限度地提高成功率和我们研究的数据，即体内抗肿瘤反应和先进的单分子显微镜。(Iv)用于流式细胞术和荧光显微镜的新探针和配体的表征和测试。(V)小分子文库的筛选，例如使用FRET探针作为读出。该流式细胞仪具有以下两个关键功能：(I)五条激光线路(包括UV)，用于通过光谱分析检测多达的荧光团。(Ii)用于细胞和组织样品的中等吞吐量实验的96孔板取样器，以及小分子抑制剂库。

Our studies seek to understand how the spatial organization and assembly dynamics of plasma membrane signaling pathways control T cell activation and functions. The research is based on our findings that plasma membrane proteins are localized in nanometer-sized domains. Our studies revealed that T cell activation is dynamically controlled by the spatial relationships of distinct nanodomains, the assembly of signaling complexes within them and translocation of signaling molecules between them. We study the underlying molecular mechanisms using multidisciplinary strategies that combine single molecule microscopy with biochemical, biophysical, and cell biological approaches. Specifically, we are interested in T cell receptor (TCR) and the inhibitory program cell death protein (PD-1) pathways during T cell activation and anti-tumor responses. Our ultimate goal is to identify novel control principles in T cell signaling pathways and use them to modify T cell responses for future immunotherapies.We request a flow cytometer with spectral analyzer and autosampler, which is essential to our newly established laboratory at the Albert-Ludwigs-Universität Freiburg and for realizing our current and future research goals. The requested flow cytometer is essential for multiple purposes, including: (i) Studying immune cell diversity during immune responses and in the tumor environment.(ii) Analyses of T cell development, differentiation and activity states. (iii) Quality control of primary and genetically modified cells to maximize success rates and data of our studies, i.e. in vivo anti-tumor responses and advanced single molecule microscopy.(iv) Characterization and testing of new probes and ligands for flow cytometry and fluorescence microscopy.(v) Screening of small molecule libraries, e.g. with FRET probes as read-outs. The flow cytometer has the following two key capabilities:(i) Five laser lines (including UV) for detection of up to 64 fluorophores through spectral analyses.(ii) A 96-wellplate sampler for medium-throughput experiments of cellular and tissue samples, as well as small molecule inhibitor libraries.