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. 2017 Nov 22;7:16044. doi: 10.1038/s41598-017-15907-3

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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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Schematic illustration of the integrated fluorescence analysis. Grey area shows a cell. Region 1 (solid ellipse) encompasses a whole cell. Region 2 (broken ellipse) encompasses region 1. Neither region (1 or 2) contains cells or fluorescent obstacles other than the cell of interest. I _cell is the integrated fluorescence over a whole cell. I ₁ and I ₂ are the integrated fluorescence values within region 1 and region 2, respectively. S ₁ and S ₂ are the areas of region 1 and region 2, respectively.