PMID- 28651016 OWN - NLM STAT- MEDLINE DCOM- 20170918 LR - 20191210 IS - 1932-6203 (Electronic) IS - 1932-6203 (Linking) VI - 12 IP - 6 DP - 2017 TI - Enhanced light microscopy visualization of virus particles from Zika virus to filamentous ebolaviruses. PG - e0179728 LID - 10.1371/journal.pone.0179728 [doi] LID - e0179728 AB - 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. FAU - Daaboul, George G AU - Daaboul GG AD - nanoView Diagnostics Inc., Boston, MA, United States of America. FAU - Freedman, David S AU - Freedman DS AD - nanoView Diagnostics Inc., Boston, MA, United States of America. FAU - Scherr, Steven M AU - Scherr SM AD - Department of Mechanical Engineering, Boston University, Boston, MA, United States of America. FAU - Carter, Erik AU - Carter E AD - Department of Microbiology, Boston University School of Medicine, Boston, MA, United States of America. FAU - Rosca, Alexandru AU - Rosca A AD - nanoView Diagnostics Inc., Boston, MA, United States of America. FAU - Bernstein, David AU - Bernstein D AD - nanoView Diagnostics Inc., Boston, MA, United States of America. FAU - Mire, Chad E AU - Mire CE AD - Galveston National Laboratory, Galveston, TX, United States of America. AD - Department of Microbiology, Galveston, TX, United States of America. AD - Immunology, University of Texas Medical Branch, Galveston, TX, United States of America. FAU - Agans, Krystle N AU - Agans KN AD - Galveston National Laboratory, Galveston, TX, United States of America. AD - Department of Microbiology, Galveston, TX, United States of America. AD - Immunology, University of Texas Medical Branch, Galveston, TX, United States of America. FAU - Hoenen, Thomas AU - Hoenen T AD - 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. AD - Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Greifswald-Isle of Riems, Germany. FAU - Geisbert, Thomas W AU - Geisbert TW AD - Galveston National Laboratory, Galveston, TX, United States of America. AD - Department of Microbiology, Galveston, TX, United States of America. AD - Immunology, University of Texas Medical Branch, Galveston, TX, United States of America. FAU - Unlu, M Selim AU - Unlu MS AD - Department of Electrical Engineering, Boston University, Boston, MA, United States of America. AD - Department of Biomedical Engineering, Boston University, Boston, MA, United States of America. AD - Physics Department, Boston University, Boston, MA, United States of America. FAU - Connor, John H AU - Connor JH AD - Department of Microbiology, Boston University School of Medicine, Boston, MA, United States of America. AD - Department of Biomedical Engineering, Boston University, Boston, MA, United States of America. LA - eng PT - Evaluation Study PT - Journal Article PT - Validation Study DEP - 20170626 PL - United States TA - PLoS One JT - PloS one JID - 101285081 SB - IM MH - Animals MH - Ebolavirus/*ultrastructure MH - Equipment Design MH - Humans MH - Microscopy, Electron, Scanning MH - Microscopy, Interference/instrumentation/*methods MH - Microscopy, Ultraviolet/instrumentation/*methods MH - Vaccinia virus/ultrastructure MH - Vesiculovirus/ultrastructure MH - Virion/*ultrastructure MH - Zika Virus/*ultrastructure PMC - PMC5484481 COIS- Competing Interests: George G. Daaboul, David S. Freedman, Alexandru Rosca and David Bernstein are employed by nanoView Diagnostics Inc. There are no patents, products in development or marketed products to declare. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors. EDAT- 2017/06/27 06:00 MHDA- 2017/09/19 06:00 PMCR- 2017/06/26 CRDT- 2017/06/27 06:00 PHST- 2017/04/07 00:00 [received] PHST- 2017/06/02 00:00 [accepted] PHST- 2017/06/27 06:00 [entrez] PHST- 2017/06/27 06:00 [pubmed] PHST- 2017/09/19 06:00 [medline] PHST- 2017/06/26 00:00 [pmc-release] AID - PONE-D-17-13654 [pii] AID - 10.1371/journal.pone.0179728 [doi] PST - epublish SO - PLoS One. 2017 Jun 26;12(6):e0179728. doi: 10.1371/journal.pone.0179728. eCollection 2017.