Abstract：
Endocytosis is a fundamental biological process that couples exocytosis to maintain the homeostasis of the plasma membrane and sustained neurotransmission. Super-resolution microscopy enables optical imaging of exocytosis and endocytosis in live cells and makes an essential contribution to understanding molecular mechanisms of endocytosis in neuronal somata and other types of cells. However, visualization of exo-endocytic events at the single vesicular level in a synapse with optical imaging remains a great challenge to reveal mechanisms governing the synaptic exo-endocytotic coupling. In this protocol, we describe the technical details of stimulated emission depletion (STED) imaging of synaptic endocytosis at the single-vesicle level, from sample preparation and microscopy calibration to data acquisition and analysis.
摘要：
胞吞作用是一个基本的生物学过程，它将胞吐结合起来以维持质膜的稳态和持续的神经传递。超分辨率显微镜可以对活细胞中的胞吐作用和内吞作用进行光学成像，并为理解神经元躯体和其他类型细胞中内吞作用的分子机制做出了重要贡献。然而,通过光学成像在突触中单个囊泡水平上的外吞事件的可视化仍然是揭示控制突触外吞-内吞耦合的机制的巨大挑战。在这个协议中,我们描述了单囊泡水平的突触内吞的受激发射耗竭（STED）成像的技术细节，从样品制备和显微镜校准到数据采集和分析。
