Measuring Viral Fusion Kinetics with Fluorescence
Author Information
Author(s): Floyd Daniel L., Harrison, Stephen C. van Oijen, Antoine M.
Primary Institution: Harvard Medical School
Hypothesis
Can we monitor the kinetics of single virus particles fusing with a target bilayer using a two-color fluorescence assay?
Conclusion
The developed assay allows for the detailed observation of viral fusion events at the single-particle level.
Supporting Evidence
- Fluorescence intensity trajectories allow for the determination of hemifusion and pore formation times.
- About 50% of particles labeled with Rh110C18 yield dequenching signals.
- 30% of SRB labeled particles provide usable signals.
- Approximately 10% of particles labeled with both dyes show both lipid mixing and pore formation signals.
Takeaway
Scientists created a special way to watch how viruses merge with cells, using colored lights to see what happens to each virus.
Methodology
An in vitro, two-color fluorescence assay was developed to monitor the kinetics of single virus particles fusing with a target bilayer.
Limitations
Prolonged imaging can bleach fluorescent labels, and synchronization of the fusion reaction can be affected by pH gradients.
Digital Object Identifier (DOI)
Want to read the original?
Access the complete publication on the publisher's website