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使用全内反射荧光(TIRF)显微镜对脂质双层中的横向迁移率和离子通道活性进行单分子成像
Journal JoVE
Biochimie
This content is Free Access.
Journal JoVE Biochimie
Single-Molecule Imaging of Lateral Mobility and Ion Channel Activity in Lipid Bilayers using Total Internal Reflection Fluorescence (TIRF) Microscopy
DOI:

08:55 min

February 17, 2023

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Chapitres

  • 00:05Introduction
  • 00:43Preparation of Lipids and Agarose Hydrogel
  • 01:33PMMA Chamber Assembly and Lipid Monolayer Formation
  • 03:06Preparation of Lipid-Coated Aqueous Droplets in Hexadecane/Silicone Oil Solution
  • 04:17Preparation and Imaging of Single Ion Channels in DIB Membranes
  • 06:49Results: Isolation of TOM-CC and Correlation of Channel Activity with Lateral Mobility of TOM-CC
  • 08:18Conclusion

Summary

Traduction automatique

该协议描述了如何使用TIRF显微镜来跟踪单个离子通道并确定它们在支持的脂质膜中的活性,从而定义侧向膜运动和通道功能之间的相互作用。它描述了如何制备膜、记录数据并分析结果。

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