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. 2019 Oct 8;10:4561. doi: 10.1038/s41467-019-12535-5

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© The Author(s) 2019

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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Fig. 2 — Multi-label FLIM. a Direct measurement of fluorescence decays obtained by sweeping gate delay time t_d for orange (O, 4.9 ns), red (R, 3.4 ns), nile red (NR, ~3.1 ns), infrared (IR, 2.3 ns) and propidium iodide (PI, 14 ns) beads. Fitted decay constants τ are given. The measured Gaussian instrument response function (IRF) is plotted in black. b Intensity image of a three-label wide-field sample of orange, nile red, and infrared beads (labels strongly overlap spatially). c Lifetime image reveals spatial distribution of the labels. Lifetime is measured by fitting the decay traces at each pixel (scale bars 10 μm)