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. 2017 Jun 7;7:2926. doi: 10.1038/s41598-017-02836-4

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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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Confocal scanning microscope images of a GUV in which two fluorophore-labeled proteins were reconstituted. (A) Detection of cyt. bo ₃ labeled with ATTO 647N. The focal plane is at the middle of the vesicle. (B) Detection of ATP-synthase labeled with ATTO 594. (C) Combined image of ATTO 594 and ATTO 647N detection. (D) An image of the top of the vesicle, which is the focal plane used for the FCS measurements. The lipid composition of the vesicle was 99% DOPC and 1% DPPE functionalized with a biotinyl head group. The GUVs were immobilized on a streptavidin-coated cover slide and the solution around the vesicle was 10 mM HEPES pH 7.4, supplemented with 10 mM NaCl and 100 mM glucose.