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PLoS One. 2017 Jun 26;12(6):e0179728. doi: 10.1371/journal.pone.0179728. eCollection 2017.

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

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nanoView Diagnostics Inc., Boston, MA, United States of America.
Department of Mechanical Engineering, Boston University, Boston, MA, United States of America.
Department of Microbiology, Boston University School of Medicine, Boston, MA, United States of America.
Galveston National Laboratory, Galveston, TX, United States of America.
Department of Microbiology, Galveston, TX, United States of America.
Immunology, University of Texas Medical Branch, Galveston, TX, United States of America.
Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rocky Mountain Laboratories, Hamilton, MT, United States of America.
Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Greifswald-Isle of Riems, Germany.
Department of Electrical Engineering, Boston University, Boston, MA, United States of America.
Department of Biomedical Engineering, Boston University, Boston, MA, United States of America.
Physics Department, Boston University, Boston, MA, United States of America.


Light microscopy is a powerful tool in the detection and analysis of parasites, fungi, and prokaryotes, 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. Virions 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.

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