Imaging/High content screening system

我们申请了基于微量滴定板的明场和荧光成像/高含量筛选系统。该系统允许快速，部分自动化的成像，定量和分析的细胞特征，以及小生物体，如线虫秀丽隐杆线虫。我们的团队研究了粘附G蛋白偶联受体（aGPCR）的相关性，以了解它们如何将信号转化为不同生物过程中的功能。粘附GPCR是一类有趣的GPCR，在健康和疾病中具有不同的作用。尽管它们具有明显的（病理）生理重要性，但关于它们的激活、信号及其生理后果的细节仍然只有很少的了解。我们的工作分析了这些独特的GPCR的分子机制，旨在阐明它们如何在生理过程中整合信号的问题。为了了解受体如何控制生理功能，我们正在阐明aGPCR在不同背景下的作用：在细胞水平上，在组织和器官中，以及在整个生物体中，如线虫线虫。因此，我们的分析强烈依赖于aGPCR的结构-功能分析。为此，我们首先研究了aGPCR缺失对细胞和线虫C的影响。优雅这主要通过表型分析来实现。随后，研究了不同修饰或天然存在的受体变体对这些表型的影响。在第二种方法中，我们的目标是鉴定能够调节aGPCR的活化/活性的分子。对于表型和结构-功能分析以及分子筛选，需要结合大型设置和大量样品的多种测定。因此，成像/高含量筛选系统是必不可少的。所要求的系统是灵活和通用的，足以在细胞以及线虫中执行所有非常不同的应用。它可以测量不同的细胞参数，如细胞大小，生长，活力，增殖，凋亡和迁移。同时，它能够成像和量化C。elegans个人在整个井和非常快。除了这些相对低分辨率的应用外，该系统还提供了放大单细胞以可视化荧光以及受体表达的可能性。该系统将大大推进我们的实验组合，并允许新的进展，如化合物筛选和大型结构功能分析，以深入了解aGPCR功能。

We apply for a microtiter plate-based brightfield and fluorescence imaging/high content screening system. This system allows for fast, partially automated imaging, quantification and analysis of cellular characteristics as well as small organisms such as the nematode Caenorhabditis elegans. Our group studies the relevance of Adhesion G protein-coupled receptors (aGPCRs) to understand how they translate signals into function in distinct biological processes. Adhesion GPCRs are an intriguing class of GPCR with diverse roles in health and disease. Despite their obvious (patho-)physiological importance, the details on their activation, signals and the physiological consequences thereof are still only sparsely understood. Our work analyses the molecular mechanisms of these unique GPCRs and aims to shed light on the question how they integrate signals in physiological processes. To understand how the receptors control physiological functions, we are elucidating the roles of aGPCRs in different contexts: on a cellular level, in tissues and organs, and in entire organisms such as the nematode Caenorhabditis elegans. Thereby, our analyses strongly depend on structure-function analyses of aGPCRs. For this purpose, we first investigate the effects the absence of an aGPCR has on cells and on the nematode C. elegans. This is mainly achieved by phenotyping assays. Subsequently, the effects of different modified or naturally occurring receptor variants on these phenotypes are investigated. In a second approach, we aim to identify molecules able to modulate the activation/activity of aGPCR. For both, phenotyping and structure-function analyses as well as molecule screenings, multiple assays combined with large set ups and numerous samples are required. Thus, an imaging/high content screening system is indispensable. The requested system is flexible and versatile enough to perform all the very different applications in cells as well as in the nematode. It can measure different cellular parameters such as cell size, growth, viability, proliferation, apoptosis, and migration. At the same time, it is able to image and quantify C. elegans individuals in whole-wells and extremely fast. In addition to these relatively low-resolution applications, the system offers the possibility to zoom in on single cells to visualize fluorescence and thus, receptor expression. The system will greatly advance our experimental portfolio and allow for novel advances such as compound screens and large structure-function analyses to gain insights into aGPCR functions.