Stimulated emission depletion microscope

由于超分辨率成像的进步，了解纳米尺度下基本细胞过程中所涉及的因素的动态和时空组织已经成为可能。在拥挤的细胞环境中使用标准衍射限制成像技术难以成像分子过程，这阻碍了进展，但商业显微镜的可用性使超分辨率技术可用于更广泛的生物界。此外，最近实施的多色超分辨率成像技术现在允许我们在完整的活细胞中跟踪这些过程，避免由于固定而产生的伪影。我们在这里建议购买一个商业受激发射损耗显微镜（STED），以解决这些关键问题的高尔基体生物学，细胞骨架在神经元功能中的作用，细胞信号网络，抗原呈递，T细胞活化和囊泡回收在神经元突触前。纳米显微镜将需要多色3D STED功能，以深入了解复杂的细胞结构。此外，我们将实施多色活细胞STED成像作为常规工具，以探索纳米级的细胞内动态过程。

Understanding the dynamics and the spatio-temporal organization of factors involved in essential cellular processes at the nanoscale has become possible thanks to the advancement of super-resolution imaging. Progress had been hampered by the difficulty to image molecular processes in the crowded cellular environment using standard diffraction limited imaging techniques, but the availability of commercial microscopes is making super-resolution techniques available to the broader biology community. Additionally, the recent implementation of multi-colour super-resolution imaging techniques now allows us to follow these processes in an intact living cell, avoiding artifacts due to fixation. We are here proposing the purchase of a commercial stimulated emission depletion microscope (STED) to address some of these key questions about Golgi biology, the role of the cytoskeleton in neuronal function, cellular signaling networks, antigen presentation, T cell activation and vesicle recycling at the neuronal presynapse. The nanoscope will require multi-color 3D STED capabilities to gain insights into the complex cellular architecture. Additionally, we will implement multi-color live-cell STED imaging as a routine tool to explore intra-cellular dynamic processes at the nanoscale.