Shaoqin Hu1 · Zhenli Xie1 · Bianbian Wang1 · Yang Chen1 · Zexin Jing1 · Ying Hao1 · Jingyu Yao1 · Xuanang Wu1 · Jingxiao Huo1 · Anqi Wei1 · Yuhao Qin3 · Nan Dong1 · Chaowen Zheng1 · Qian Song1 · Jiangang Long1 · Xinjiang Kang2,3 · Changhe Wang1,3 · Huadong Xu11 Neuroscience Research Center, Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Department of Neurology, the First Afliated Hospital, Core Facilities Sharing Platform, Xi’an Jiaotong University, Xi’an 710049, China
2 College of Life Sciences, Liaocheng University, Liaocheng 252059, China
3 Key Laboratory of Medical Electrophysiology, Ministry of Education of China, Collaborative Innovation Center for Prevention and Treatment of Cardiovascular Disease, and the Institute of Cardiovascular Research, Southwest Medical University, Luzhou 646000, China
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.
Keywords
Endocytosis; STED imaging; Synaptic vesicle; Single-vesicle level
