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. 2019 May 29;9:8051. doi: 10.1038/s41598-019-44537-0

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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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Live imaging and bioinformatic data acquisition. (A) Diagram of the Fucci System. (B) Experimental approach. (C) Time-lapse images of a representative FUCCI-H2B colony in naive ground state conditions growing for 45 h. Fluorescent images are merged compositions of the three channels corresponding to hCd1-mCherry (G1), hGeminin-mVenus (SG2M) and H2B-mCerulean (nuclear marker). (D) Schematic of the different steps performed to acquire and analyze single cell live imaging data. (E) hCd1-mCherry and hGeminin-mVenus fluorescence dynamics for a representative cell over time. CC-L was defined as the length of time since a cell was born and up to its division, deduced by mitotic events analyzed with the H2B nuclear marker. G1-L was defined as the time since a cell was born and up to the local maximum of the hCd1-mCherry fluorescence, while SG2M-L was calculated as the difference between CC-L and G1-L. (F) Representative dendrogram reconstructed for a lineage of cells growing in differentiating conditions.