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. 2018 Sep 14;8:14067. doi: 10.1038/s41598-018-32301-9

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© The Author(s) 2018

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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Fluorescence lifetime and fluorescence anisotropy images of a single exosome. Panel A shows a cell from the HT-29 cell line, cultured at AmB concentration of 10 µg/ml. Panel B shows an image of a single exosome vesicle, selected in panel A (marked with a white circle) and recorded with higher resolution. Panel C shows the same exosome imaged based on fluorescence anisotropy data. The colour code for panels A and B as in Fig. 2. The relatively short fluorescence lifetimes of AmB in the exosome (panel B) is indicative of the formation of small aggregated forms of the drug in the lipid phase. The relatively high fluorescence anisotropy values at the left- and right-hand of the image of a vesicle cross-section (panel C) are consistent with the perpendicular orientation of AmB molecules with respect to the membrane plane.