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. 2017 Oct 23;7:13772. doi: 10.1038/s41598-017-14226-x

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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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A sequential injection strategy rescues TMRE intensity from the decrease caused by simultaneous injection with 2-NBDG. TMRE and 2-NBDG kinetic imaging was performed on murine dorsal window chambers after they received one of three distinct injection strategies: (1) TMRE alone, (2) 2-NBDG alone, (3) TMRE followed by 2-NBDG with a 10–15 min delay. (a) Representative TMRE uptake time course images for the sequential injection strategy and independent TMRE injection. (b) Representative 2-NBDG uptake time course images for the sequential injection strategy and independent 2-NBDG injection. (c) Mean uptake kinetics of TMRE for each injection strategy. (d) Mean uptake kinetics of 2-NBDG for each injection strategy. NS = not significant. N = 5–6 mice/group. Comparison of mean kinetic curves across animal groups was performed with a two-way Analysis of Variance (ANOVA) test using the MATLAB (Mathworks, USA) statistics toolbox.