Automatisiertes Multiplex-Live-Cell-Imaging-System

在很长一段时间内实时定量评估细胞行为通常提供对所评估的细胞过程的生理相关性的惊人洞察力，并有力地补充了详细的生化和高通量组学研究。直到最近，这些实验的意义往往受到成像资源的时间限制、成像过程中细胞扰动造成的实验伪影以及仅供专家用户使用的复杂分析软件的限制。然而，自动化活细胞成像技术的最新发展使我们能够在不受干扰的培养条件下，以单细胞分辨率实时检测细胞代谢和氧化还原行为、细胞分化和运动、细胞增殖、细胞周期进展、类器官和类球形形成以及细胞死亡。这些现代的多路自动活细胞成像系统与各种经过验证的无标记、彩色和荧光分析结合使用。此外，硬件设置通常具有足够的通用性，一旦用于视觉或基于荧光的监测的分析可用，就允许添加更多的测量参数。该设备旨在作为科隆大学分子生物科学中心(COMB)的中心仪器使用。该研究所由遗传学、生物化学、生物物理学和植物学研究所的工作组组成，以跨学科的方式研究细胞生理学、新陈代谢和生命周期。Comb的研究小组主要使用培养细胞、原代细胞、器官培养和细胞分化系统的模型来研究“细胞生物化学”领域的不同主题。对于这项研究，需要大量基于细胞的分析，并具有相当大的吞吐量。本提案中应用的自动化多路活细胞成像系统允许进行广泛的表型研究。此外，它允许对大量样品进行独立且高度可重复性的并行延时分析(例如6 x 96孔板)，从而满足梳子内基于细胞的分析的高要求。因此，收购这一现代化的多路自动活细胞成像系统将是COMB和科隆校区研究小组开展的许多项目和合作努力的重要资产。

Assessing cell behaviour in real time quantitatively over long time periods often provides striking insights into the physiological relevance of assessed cellular processes and strongly complements detailed biochemical and high throughput omics studies. Until recently, the significance of these experiments was often restricted by time-limited access to imaging resources, experimental artefacts caused by perturbances of cells in the process of imaging and complex analysis software solely dedicated to expert users. However, recent developments in automated live cell imaging techniques enable examining cellular metabolic and redox behaviour, cellular differentiation and motility, cell proliferation, cell cycle progression, organoid and spheroid formation and cell death in real time with single cell resolution under unperturbed culture conditions. These modern Automated Multiplex Live Cell Imaging Systems are used in conjunction with various validated label-free, colour and fluorometric assays. Additionally, the hardware setups usually are sufficiently versatile to allow adding further measurement parameters as soon as assays for their visual or fluorescence-based monitoring become available. The device is intended for use as a central instrument in the Center for Molecular Biosciences (CoMB) at the University of Cologne. The CoMB is made up of working groups from the Institutes of Genetics, Biochemistry, Biophysics and Botany to investigate cellular physiology, metabolism and life cycles in an interdisciplinary manner. Research groups at the CoMB work on different topics in the area of “cellular biochemistry” using mainly models of cultured cells, primary cells, organoid cultures and systems for cell differentiation. For this research a large number of cell-based assays with a substantial throughput are required. The Automated Multiplex Live Cell Imaging System applied for in this proposal allows a wide variety of phenotypic studies. Moreover, it allows independent and highly reproducible time-delayed analyses of high numbers of samples in parallel (e.g. 6 x 96 well plates) and thus meeting the high demand of cell-based assays within the CoMB. Therefore, the acquisition of this modern Automated Multiplex Live Cell Imaging System will be a vital asset to many projects and cooperative endeavours carried out by the research groups at the CoMB and the Cologne campus.