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单脂蛋白的SNARE介导融合在一个微流控流细胞拴支持双分子层通过偏光显微镜TIRF监控
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
SNARE-mediated Fusion of Single Proteoliposomes with Tethered Supported Bilayers in a Microfluidic Flow Cell Monitored by Polarized TIRF Microscopy
DOI:

10:58 min

August 24, 2016

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Chapters

  • 00:05Title
  • 01:01Preparation of a PDMS Block to Form the Microfluidic Channel
  • 03:17Formation of the Tethered Supported Bilayer
  • 06:18Checking Bilayer Fluidity
  • 07:06Introducing v-SUVs into the Microfluidic Channels
  • 07:59Observing the Fusion of Single v-SUVs and the SBL
  • 08:42Results: Docking and Fusion Events Described by Lipid and Content Release
  • 10:26Conclusion

Summary

Automatic Translation

在这里,我们提出了一个协议来检测使用偏振TIRFM脂质体和微流体通道支持双层之间的单,SNARE介导的融合事件,与单分子敏感性和〜15毫秒的时间分辨率。脂质和可溶性货物放行可同时检测。脂质体大小,脂质扩散率,和融合孔性质被测量。

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