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. 2017 Aug 11;7:7983. doi: 10.1038/s41598-017-08168-7

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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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Detection of the emission spectra of the fast and slow components of ICG dye (1 µM) in the heart. (A) Superimposition of spectra recorded at λ_ex = 660 nm; total fluorescence was normalized to 100%. Four separate spectra obtained at rest (solid) and at the maximum amplitude of the AP (dash): two for the fast component (F_f1 and F_f2; red and dark red) and two for the slow component (F_s1 and F_s2; blue and dark blue) are presented alongside the calculated total OS (F and F + ΔF, green). The spectrum peaks on an expanded fluorescence scale are shown in the insets. (B) Superimposition of fractional fluorescence signals at λ_ex/em = 660/800 nm. Experimentally detected with a fast EMCCD camera the total voltage-sensitive OS (ΔF_exp; violet), from the total OS calculated fast (ΔF_f; red) and slow (ΔF_s; blue) components, and recalculated OS (ΔF, the sum of ΔF_f and ΔF_s; green). (C–E) The ICG fluorescence obtained at λ_ex = 660 and λ_em from 720 nm to 905 nm for the total fractional fluorescence of OS (ΔF; violet) and for both components: ΔF_f (red) and ΔF_s (blue). The data shown in all panels were obtained in a single experiment.