Analysis and Sorting of Individual HSV-1 Particles by Flow Virometry
| Autor: | Nabil El Bilali, Bita Khadivjam, Roger Lippé |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification medicine.diagnostic_test biology viruses Virulence HSL and HSV Computational biology Viral tegument medicine.disease_cause Flow cytometry 03 medical and health sciences 030104 developmental biology 0302 clinical medicine Herpes simplex virus Capsid chemistry 030220 oncology & carcinogenesis medicine biology.protein Antibody Glycoprotein |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493998135 |
| DOI: | 10.1007/978-1-4939-9814-2_16 |
| Popis: | Flow cytometry has been instrumental in characterizing normal and infected cells. However, until recently, it was not possible to use such an approach to analyze small entities such as bacteria, let alone viruses, owing to the 0.5 μm resolution of most instruments. To circumvent this limitation, some laboratories decorate pathogens with antibodies or nanoparticles. Our laboratory instead exploits an alternative approach that relies on the staining of internal viral constituents with permeable SYTO dyes or the fluorescent tagging of individual viral proteinaceous components, whether capsid, tegument or glycoproteins. This opens up a range of new research avenues and, for example, enabled us to characterize individual herpes simplex virus type 1 particles, discern their different subpopulations, measure the heterogeneity of mature virions in terms of protein content, sort these viral particles with >90% purity and, for the first time, directly address the impact of this heterogeneity on viral fitness. This approach, coined flow virometry or nanoscale flow cytometry, allows for the study of a wide variety of pathogens with high statistical significance and the potential discovery of novel virulence factors. |
| Databáze: | OpenAIRE |
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