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. 2017 Jun 19;7:3779. doi: 10.1038/s41598-017-03610-2

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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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Staining variation between multiple identical samples prepared separately. (a) Histograms of a single sample of H9 cells split into three and stained separately following identical protocol. (b) Second replicate using a second sample split into three. (c) Cells were incubated with the indicated concentrations of cisplatin (Pt196) for 5 min. Signal distributions for the Pt196 channel as well as the measured contributions of the cisplatin Pt196 reagent at the relative concentrations to the Pt194 and Pt195 channels are shown. (d) Cells were incubated with 100 nM cisplatin (Pt196) for the indicated time periods. Signal distributions for Pt196 are shown. (e) Eight PFA-fixed H9 cell samples labeled with eight combinations of the Pt194, Pt195 and Pt196 cisplatin regents, pooled, and run as a single sample producing eight distinct populations.