STED Imaging of Vesicular Endocytosis in the Synapse.
Shaoqin HuZhenli XieBianbian WangYang ChenZexin JingYing HaoJingyu YaoXuanang WuJingxiao HuoAnqi WeiYuhao QinNan DongChaowen ZhengQian SongJiangang LongXinjiang KangChanghe WangHuadong XuPublished in: Neuroscience bulletin (2024)
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.
Keyphrases
- high resolution
- induced apoptosis
- high speed
- cell cycle arrest
- randomized controlled trial
- mass spectrometry
- gene expression
- single molecule
- electronic health record
- signaling pathway
- oxidative stress
- endoplasmic reticulum stress
- brain injury
- tandem mass spectrometry
- big data
- deep learning
- low cost
- solid state
- simultaneous determination