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. 2017 Nov 15;7:15609. doi: 10.1038/s41598-017-15411-8

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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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Measured EEM of Fluorescein and TAMRA solutions at 2 $μ$ M concentration in PBS, (A) and (B) respectively. Donor, Acceptor, and FRET components used for linear unmixing were generated from the normalized excitation and emission spectra as shown in (C), (D), and (E), respectively. Panel F shows the EEM of a 1:1 mixture of the Fluorescein and TAMRA solutions with the relative magnitudes of the fitting components superimposed (Donor: 55.2%, Acceptor: 45.8%, FRET: −1.0%)