ArticleFrontiers in synaptic neuroscience2016
Flash-and-Freeze: Coordinating Optogenetic Stimulation with Rapid Freezing to Visualize Membrane Dynamics at Synapses with Millisecond Resolution.
Article in Frontiers in synaptic neuroscience, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 30 citations in OpenAlex.
- Membrane Merger Dynamics in Exocytosis and Endocytosis.Advances in experimental medicine and biology · 2026Review
- Cryo-ET of actin cytoskeleton and membrane structure in lamellipodia formation using optogenetics.iScience · 2025Article
- Interpretation of presynaptic phenotypes of synaptic plasticity in terms of a two-step priming process.The Journal of general physiology · 2024Article
- Coming of Age: Cryo-Electron Tomography as a Versatile Tool to Generate High-Resolution Structures at Cellular/Biological Interfaces.International journal of molecular sciences · 2021Review
- A Compartmentalized Neuronal Cell-Culture Platform Compatible With Cryo-Fixation by High-Pressure Freezing for Ultrastructural Imaging.Frontiers in neuroscience · 2021Article
- Synaptic vesicles transiently dock to refill release sites.Nature neuroscience · 2020Article
- In situ Microfluidic Cryofixation for Cryo Focused Ion Beam Milling and Cryo Electron Tomography.Scientific reports · 2019Article
- Synaptic Vesicle Endocytosis in Different Model Systems.Frontiers in cellular neuroscience · 2018Review
- Flash-and-Freeze: A Novel Technique to Capture Membrane Dynamics with Electron Microscopy.Journal of visualized experiments : JoVE · 2017Article
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Authors and funding
1 author at 1 institution in 1 country.
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Abstract
Electron microscopy depicts subcellular structures at synapses exquisitely but only captures static images. To visualize membrane dynamics, we have developed a novel technique, called flash-and-freeze, which induces neuronal activity with a flash of light and captures the membrane dynamics by rapid freezing. For characterizing membrane movements during synaptic transmission, a light-sensitive cation channel, channelrhodopsin, is heterologously expressed in mouse hippocampal neurons or in Caenorhabditis elegans motor neurons. A brief pulse of blue light activates channelrhodopsin and induces an action potential, leading to synaptic transmission. Following the light stimulation, neurons are frozen at different time intervals ranging from 10 ms to 20 s. Electron micrographs are then acquired from each time point to visualize the morphological changes. Using this approach, we have characterized a novel form of endocytosis, ultrafast endocytosis, which rapidly removes excess membrane added to the surface during neurotransmission. The flash-and-freeze approach can be adapted to study other cellular phenomena that can be induced by light-sensitive genetic or pharmacological tools.
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