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Analysis and Sorting of Individual HSV-1 Particles by Flow Virometry
Bita Khadivjam1, Nabil El Bilali1, Roger Lippé2
1Department of Pathology and Cell Biology, University of Montreal, Montreal, QC, Canada.
Methods in Molecular Biology (Clifton, N.J.)
|October 17, 2019
Summary
Flow virometry enables the analysis of individual virus particles by staining internal components. This breakthrough allows researchers to study viral heterogeneity and its impact on fitness for the first time.
Area of Science:
- Virology
- Biotechnology
- Cell Biology
Background:
- Flow cytometry is a powerful tool for cell analysis but limited by resolution for small pathogens like viruses.
- Previous methods involved modifying pathogens, limiting direct analysis.
- Analyzing individual viral particles was previously not feasible.
Purpose of the Study:
- To develop a novel method for analyzing individual virus particles using flow cytometry.
- To overcome the resolution limitations of conventional flow cytometry for viral studies.
- To characterize viral heterogeneity and its impact on viral fitness.
Main Methods:
- Developed "flow virometry" or "nanoscale flow cytometry" by staining internal viral components with SYTO dyes.
- Alternatively, fluorescently tagged individual viral proteinaceous components (capsid, tegument, glycoproteins).
- Applied the technique to herpes simplex virus type 1 (HSV-1) for particle characterization and sorting.
Main Results:
- Successfully characterized individual HSV-1 particles and identified distinct subpopulations.
- Quantified heterogeneity in protein content among mature virions.
- Achieved >90% purity in sorting viral particles based on their characteristics.
- Enabled direct assessment of heterogeneity's impact on viral fitness.
Conclusions:
- Flow virometry offers a new approach for studying a wide range of pathogens at the single-particle level.
- This technique allows for high statistical significance in pathogen analysis.
- Potential for discovering novel virulence factors by understanding viral heterogeneity.
