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Method for Measurement of Viral Fusion Kinetics at the Single Particle Level
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Biologia
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JoVE Journal Biologia
Method for Measurement of Viral Fusion Kinetics at the Single Particle Level

Method for Measurement of Viral Fusion Kinetics at the Single Particle Level

DOI:
10.3791/1484-v

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14:59 min

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September 07, 2009

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, ,
1Department of Biological Chemistry and Molecular Pharmacology,Harvard Medical School, 2Howard Hughes Medical Institute,Harvard Medical School

Capitoli

  • 00:04Titolo
  • 00:49Introduction
  • 01:12Preparing Dextran-Functionalized Coverslips
  • 04:29Preparing the Liposomes
  • 06:41Setting Up the Microfluidic Flow Cell
  • 08:28Labeling the Virus Particles
  • 10:16Observing Viral Fusion Kinetics Under the Microscope
  • 13:10Viral Hemifusion and Pore Formation Data: Microscope Footage, Fluorescence Intensity Trajectories, and Lag Times for Viral Hemifusion and Pore Formation
  • 14:30Conclusion

Summary

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We present an in vitro, two-color fluorescence assay to visualize the fusion of single virus particles with a fluid target bilayer. By labeling viral particles with fluorophores that differentially stain the viral membrane and its interior, we are able to monitor the kinetics of hemifusion and pore formation.

Tags

Viral Fusion KineticsSingle Particle LevelMembrane FusionFusion AssayFluorescence AssayFluorphore StainingGanglioside-containing Lipid BilayerCCD CameraPH-dependent FusionHemifusionPore Formation

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