Enhanced light microscopy visualization of virus particles from Zika virus to filamentous ebolaviruses

George G. Daaboul, David S. Freedman, Steven M. Scherr, Erik Carter, Alexandru Rosca, David Bernstein, Chad Mire, Krystle N. Agans, Thomas Hoenen, Thomas Geisbert, M. Selim Ünlü, John H. Connor

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Light microscopy is a powerful tool in the detection and analysis of parasites, fungi, and pro-karyotes, but has been challenging to use for the detection of individual virus particles. Unlabeled virus particles are too small to be visualized using standard visible light microscopy. Characterization of virus particles is typically performed using higher resolution approaches such as electron microscopy or atomic force microscopy. These approaches require purification of virions away from their normal millieu, requiring significant levels of expertise, and can only enumerate small numbers of particles per field of view. Here, we utilize a visible light imaging approach called Single Particle Interferometric Reflectance Imaging Sensor (SP-IRIS) that allows automated counting and sizing of thousands of individual virions. Viri-ons are captured directly from complex solutions onto a silicon chip and then detected using a reflectance interference imaging modality. We show that the use of different imaging wavelengths allows the visualization of a multitude of virus particles. Using Violet/UV illumination, the SP-IRIS technique is able to detect individual flavivirus particles (~40 nm), while green light illumination is capable of identifying and discriminating between vesicular stomatitis virus and vaccinia virus (~360 nm). Strikingly, the technology allows the clear identification of filamentous infectious ebolavirus particles and virus-like particles. The ability to differentiate and quantify unlabeled virus particles extends the usefulness of traditional light microscopy and can be embodied in a straightforward benchtop approach allowing widespread applications ranging from rapid detection in biological fluids to analysis of virus-like particles for vaccine development and production.

Original languageEnglish (US)
Article numbere0179728
JournalPLoS One
Volume12
Issue number6
DOIs
StatePublished - Jun 1 2017

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Zika virus
Ebolavirus
Viruses
virion
Virion
Optical microscopy
light microscopy
Microscopy
Visualization
Light
image analysis
Imaging techniques
reflectance
sensors (equipment)
Lighting
lighting
virus-like particle vaccines
Virus-Like Particle Vaccines
Vesiculovirus
Flavivirus

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Daaboul, G. G., Freedman, D. S., Scherr, S. M., Carter, E., Rosca, A., Bernstein, D., ... Connor, J. H. (2017). Enhanced light microscopy visualization of virus particles from Zika virus to filamentous ebolaviruses. PLoS One, 12(6), [e0179728]. https://doi.org/10.1371/journal.pone.0179728

Enhanced light microscopy visualization of virus particles from Zika virus to filamentous ebolaviruses. / Daaboul, George G.; Freedman, David S.; Scherr, Steven M.; Carter, Erik; Rosca, Alexandru; Bernstein, David; Mire, Chad; Agans, Krystle N.; Hoenen, Thomas; Geisbert, Thomas; Selim Ünlü, M.; Connor, John H.

In: PLoS One, Vol. 12, No. 6, e0179728, 01.06.2017.

Research output: Contribution to journalArticle

Daaboul, GG, Freedman, DS, Scherr, SM, Carter, E, Rosca, A, Bernstein, D, Mire, C, Agans, KN, Hoenen, T, Geisbert, T, Selim Ünlü, M & Connor, JH 2017, 'Enhanced light microscopy visualization of virus particles from Zika virus to filamentous ebolaviruses', PLoS One, vol. 12, no. 6, e0179728. https://doi.org/10.1371/journal.pone.0179728
Daaboul GG, Freedman DS, Scherr SM, Carter E, Rosca A, Bernstein D et al. Enhanced light microscopy visualization of virus particles from Zika virus to filamentous ebolaviruses. PLoS One. 2017 Jun 1;12(6). e0179728. https://doi.org/10.1371/journal.pone.0179728
Daaboul, George G. ; Freedman, David S. ; Scherr, Steven M. ; Carter, Erik ; Rosca, Alexandru ; Bernstein, David ; Mire, Chad ; Agans, Krystle N. ; Hoenen, Thomas ; Geisbert, Thomas ; Selim Ünlü, M. ; Connor, John H. / Enhanced light microscopy visualization of virus particles from Zika virus to filamentous ebolaviruses. In: PLoS One. 2017 ; Vol. 12, No. 6.
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