Laser-Scanning Mikroskop mit konfokaler Optik

研究了植物茎尖分生组织中干细胞稳态的调控机制。由于干细胞群体本身以及为分生组织完整性提供组织线索的细胞经历了永久性的更新，我们需要将我们的研究从组织学切片提供的静态二维水平扩展到时间推移共聚焦显微镜实现的动态四维水平。我们开发了携带多色荧光报告基因的植物系，这将使我们能够同时看到茎尖分生组织的所有主要结构域。由于报告基因以细胞核为目标，因此可以量化给定区域内的细胞数量。此外，我们正在开发强大的标记来评估细胞分裂活性，并与细胞身份标记一起，这些工具将使我们对拟南芥顶端干细胞生态位的细胞行为有前所未有的了解。对于这些实验，我们需要使用一种多功能的直立共聚焦显微镜，该显微镜能够解析光谱信息，因为每次实验同时检测到大量标记物，时间长达数小时。此外，为了获得定量信息，我们将需要相当程度的成像自动化。到目前为止，我们的机构还没有满足这些要求的仪器，现有的仪器被高度使用，排除了这些类型的分析。

We study the regulatory mechanisms underlying plant stem cell homeostasis at the shoot apical meristem. Since the stem cell population itself as well as the cells providing the organizational cues for meristem integrity undergo permanent turnover, we need to expand our studies from the static 2D level afforded by histological sections to the dynamic 4D level achieved by time lapse confocal microscopy. We have developed plant lines carrying multi-color fluorescent reporters, which will enable us to visualize all major domains within the shoot apical meristem simultaneously. Since the reporters are targeted to the nucleus, it will be possible to quantify the cell number within a given domain. In addition we are developing powerful markers to assess cell division activity and together with cell identity markers, these tools will give us unprecedented insight into cell behavior in the apical stem cell niche of Arabidopsis. For these experiments we need access to a versatile upright confocal microscope able to resolve spectral information due to the multitude of simultaneously detected markers for many hours for each experiment. Furthermore, to acquire quantitative information, we will need a substantial level of automation for imaging. So far, an instrument that meets these requirements does not exist at our institution and the available instruments are highly used precluding these types of analysis.