Journal
/
Biochemistry
/
Single-Molecule Imaging of Lateral Mobility and Ion Channel Activity in Lipid Bilayers using Total Internal Reflection Fluorescence (TIRF) Microscopy
JoVE Journal
Biochemistry
This content is Free Access.
JoVE Journal Biochemistry
Single-Molecule Imaging of Lateral Mobility and Ion Channel Activity in Lipid Bilayers using Total Internal Reflection Fluorescence (TIRF) Microscopy

Single-Molecule Imaging of Lateral Mobility and Ion Channel Activity in Lipid Bilayers using Total Internal Reflection Fluorescence (TIRF) Microscopy

DOI:
10.3791/64970-v

•

08:55 min

•

February 17, 2023

•

,
1Biophysics Department, Institute of Biomaterials and Biomolecular Systems,University of Stuttgart

Chapters

  • 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

Automatic Translation

Automatic Translation

This protocol describes how to use TIRF microscopy to track individual ion channels and determine their activity in supported lipid membranes, thereby defining the interplay between lateral membrane movement and channel function. It describes how to prepare the membranes, record the data, and analyze the results.

Tags

Keywords: Single-molecule ImagingLateral MobilityIon Channel ActivityLipid BilayersTotal Internal Reflection Fluorescence (TIRF) MicroscopyMembrane ProteinsDPhPCHexadecaneSilicone OilAgarose HydrogelPMMA Chamber

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/64970?article_type=v

Detergent-free Ultrafast Reconstitution of Membrane Proteins into Lipid Bilayers Using Fusogenic Complementary-charged Proteoliposomes.
check_url/64970?article_type=v

In Vitro Reconstitution of Self-Organizing Protein Patterns on Supported Lipid Bilayers
check_url/64970?article_type=v

Dissipative Microgravimetry to Study the Binding Dynamics of the Phospholipid Binding Protein Annexin A2 to Solid-supported Lipid Bilayers Using a Quartz Resonator
check_url/64970?article_type=v

Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy
check_url/64970?article_type=v

Single-Molecule Tracking Microscopy – A Tool for Determining the Diffusive States of Cytosolic Molecules
check_url/64970?article_type=v

Production of Dynein and Kinesin Motor Ensembles on DNA Origami Nanostructures for Single Molecule Observation
check_url/64970?article_type=v

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip
check_url/64970?article_type=v

Conventional BODIPY Conjugates for Live-Cell Super-Resolution Microscopy and Single-Molecule Tracking
check_url/64970?article_type=v

Single Cell Analysis Of Transcriptionally Active Alleles By Single Molecule FISH
check_url/64970?article_type=v

Single Molecule Fluorescence Energy Transfer Study of Ribosome Protein Synthesis

Read Article