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Single-Molecule Diffusion and Assembly on Polymer-Crowded Lipid Membranes
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Biochemistry
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JoVE Journal Biochemistry
Single-Molecule Diffusion and Assembly on Polymer-Crowded Lipid Membranes

Single-Molecule Diffusion and Assembly on Polymer-Crowded Lipid Membranes

DOI:
10.3791/64243-v

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10:43 min

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July 19, 2022

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, , , , ,
1Department of Chemical Engineering,Indian Institute of Science, 2Holonyak Micro & Nanotechnology Lab,University of Illinois, Urbana-Champaign, 3Center for BioSystems Science and Engineering,Indian Institute of Science

Chapters

  • 00:04Introduction
  • 00:53Assembly of the Microfluidic Chamber
  • 01:41Making Supported Bilayers on Glass Substrate by Vesicle Fusion
  • 04:22Microscope Setup and Single-Particle Imaging Measurements
  • 07:26Image Acquisition for Counting Subunits in a Protein Assembly
  • 08:40Results: Analyzing the Binding of Cytolysin A (ClyA) Protein and Mobility of DNA Tracer
  • 10:10Conclusion

Summary

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Here, a protocol to perform and analyze the binding, mobility, and assembly of single molecules on artificial crowded lipid membranes using single-molecule total internal reflection fluorescence (smTIRF) microscopy is presented.

Tags

Keywords: Single-molecule ImagingPolymer-crowded Lipid MembranesSupported Lipid BilayersMembrane Biomolecular ReactionsDiffusionAssemblyPOPCDOPE-PEG(2000)Small Unilamellar VesiclesSonicationMicrofluidic Imaging Chambers

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