Konfokales Live-Imaging Mikroskop mit Spinning Disc

要在分子水平上理解细胞行为的动力学，越来越需要在受控环境(二氧化碳、温度)的活体条件下观察到单分子水平的相互作用。我们正在申请一种共聚焦显微镜，用于研究活样本中的动态细胞和细胞内过程。该仪器必须提供最先进的空间和时间分辨率。考虑到神经细胞和组织的脆弱性质，高灵敏度和尽可能低的光毒性是必不可少的。对于极其敏感的干细胞和神经元样本，必须保持稳定的环境，同时在短期和长期观察期间提供稳定的光学系统。为了跟踪细胞在时间和空间上的运动和投影，需要一个具有自动聚焦能力的全电动显微镜平台。FRET功能应该是初始配置的一部分。使用多色转基因鼠标工具需要快速和宽波长的共聚焦激发和检测，以及在可见光下同时记录的DIC能力。基于对比度的自动对焦也需要DIC。目前，Charité还没有类似的成像站，能够提供与高灵敏度神经细胞和组织培养相兼容的先进活细胞成像技术。

Understanding the dynamics of cell behavior at the molecular level increasingly requires the observation of interactions all the way to the single molecule level under in vivo conditions in a controlled environment (CO2, temperature). We are applying for a confocal microscope for the study of dynamic cellular and intracellular processes in live samples. The instrument must provide state-of-the-art spatial and temporal resolution. High sensitivity with the lowest possible phototoxicity is essential given the fragile nature of neuronal cells and tissue. A constant environment must be maintained for extremely sensitive stem cell and neuronal samples while providing for stable optics during both short and long term observations. A fully motorized microscope platform with autofocus capability is necessary for following cell movements and projections in time and space. FRET capability should be part of the initial configuration. Use of multi-color genetically modified mouse tools requires fast and broad wavelength confocal excitation and detection, together with DIC capability for simultaneous documentation in visible light. DIC is also required for autofocus based on contrast. At present there is no comparable imaging station at the Charité capable of advanced live cell imaging techniques compatible with highly sensitive neuronal cell and tissue culture.