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. 2017 Dec 19;7:17769. doi: 10.1038/s41598-017-18227-8

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Direct staining of virus. MLVeGFP viruses from chronically infected NIH 3T3 cell supernatants were labeled with (A) DiD, (B) DiI, (C) FM4–64, (D) Rhodamine-1,2-DihexadecanoylPhosphatidylethanolamine (DHPE-Rhodamine), and (E) Ceramide-BODIPY TR. Samples were then analyzed by NFC without purification. eGFP+ particles are depicted in green, background noise or non-fluorescent particles in black, dye-labeled particles in color (other than green). The distribution of total dye-labeled and unlabeled eGFP+ particles is indicated in green. Included in each panel: unstained virus (MLVeGFP), stained virus (MLVeGFP+ dye), dye alone (PBS + dye), and uninfected supernatant with dye that serves as an EV-only sample (Uninf Sup + dye).