Stimulated emission depletion microscope

由于超分辨率成像的进步，在纳米尺度上理解参与基本细胞过程的因素的动力学和时空组织成为可能。由于很难使用标准的衍射限制成像技术在拥挤的细胞环境中对分子过程进行成像，这一进展受到了阻碍，但商用显微镜的出现使超分辨率技术可以应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一个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台商用受激发射耗尽显微镜(STED)，以解决高尔基体生物学、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细胞激活和突触前囊泡回收等关键问题。纳米显微镜将需要多色3D STED能力，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，作为在纳米尺度探索细胞内动态过程的常规工具。

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.