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Flash-and-Freeze: A Novel Technique to Capture Membrane Dynamics with Electron Microscopy
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JoVE Journal 生物学
Flash-and-Freeze: A Novel Technique to Capture Membrane Dynamics with Electron Microscopy

Flash-and-Freeze: A Novel Technique to Capture Membrane Dynamics with Electron Microscopy

DOI:
10.3791/55664-v

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10:01 min

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May 01, 2017

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, ,
1Department of Cell Biology,Johns Hopkins School of Medicine, 2生化学・分子生物学科,Johns Hopkins Bloomberg School of Public Health, 3Solomon H. Snyder Department of Neuroscience,Johns Hopkins School of Medicine

章

  • 00:05標題
  • 00:52Flash-and-Freeze
  • 03:41Automated Freeze-substitution Unit-transfer
  • 06:40Sample Mounting and Imaging
  • 08:24Results: Representative Exocytosis and Endocytosis in Mouse Hippocampal Neurons
  • 08:56Conclusion

概要

自動翻訳

自動翻訳

We developed a novel technique in electron microscopy, "flash-and-freeze," that enables the visualization of membrane dynamics with ms temporal resolution. This technique combines the optogenetic stimulation of neurons with high-pressure freezing. Here, we demonstrate the procedures and describe the protocols in detail.

タグ

Electron MicroscopyMembrane DynamicsOptogeneticsSynaptic Vesicle RecyclingFlash-and-freezeHigh-pressure FreezingFree SubstitutionTime-resolved Imaging

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