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Automatic Quantitative Segmentation of Myotubes Reveals Single-cell Dynamics of S6 Kinase Activation

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dc.contributor.author Inoue, Haruki
dc.contributor.author Kunida, Katsuyuki
dc.contributor.author Matsuda, Naoki
dc.contributor.author Hoshino, Daisuke
dc.contributor.author Wada, Takumi
dc.contributor.author Imamura, Hiromi
dc.contributor.author Noji, Hiroyuki
dc.contributor.author Kuroda, Shinya
dc.date.accessioned 2019-02-12T06:08:27Z
dc.date.available 2019-02-12T06:08:27Z
dc.date.issued 2018-08-31
dc.identifier.issn 0386-7196
dc.identifier.uri http://hdl.handle.net/10061/13115
dc.description.abstract Automatic cell segmentation is a powerful method for quantifying signaling dynamics at single-cell resolution in live cell fluorescence imaging. Segmentation methods for mononuclear and round shape cells have been developed extensively. However, a segmentation method for elongated polynuclear cells, such as differentiated C2C12 myotubes, has yet to be developed. In addition, myotubes are surrounded by undifferentiated reserve cells, making it difficult to identify background regions and subsequent quantification. Here we developed an automatic quantitative segmentation method for myotubes using watershed segmentation of summed binary images and a two-component Gaussian mixture model. We used time-lapse fluorescence images of differentiated C2C12 cells stably expressing Eevee-S6K, a fluorescence resonance energy transfer (FRET) biosensor of S6 kinase (S6K). Summation of binary images enhanced the contrast between myotubes and reserve cells, permitting detection of a myotube and a myotube center. Using a myotube center instead of a nucleus, individual myotubes could be detected automatically by watershed segmentation. In addition, a background correction using the two-component Gaussian mixture model permitted automatic signal intensity quantification in individual myotubes. Thus, we provide an automatic quantitative segmentation method by combining automatic myotube detection and background correction. Furthermore, this method allowed us to quantify S6K activity in individual myotubes, demonstrating that some of the temporal properties of S6K activity such as peak time and half-life of adaptation show different dose-dependent changes of insulin between cell population and individuals. ja_JP
dc.language.iso en ja_JP
dc.publisher 日本細胞生物学会 ja_JP
dc.rights @2018 The Author(s) ja_JP
dc.subject time lapse images ja_JP
dc.subject cell segmentation ja_JP
dc.subject fluorescence resonance energy transfer ja_JP
dc.subject C2C12 ja_JP
dc.subject myotube ja_JP
dc.title Automatic Quantitative Segmentation of Myotubes Reveals Single-cell Dynamics of S6 Kinase Activation ja_JP
dc.type.nii Journal Article ja_JP
dc.contributor.transcription クニダ, カツユキ
dc.contributor.alternative 国田, 勝行
dc.identifier.fulltexturl https://doi.org/10.1247/csf.18012 ja_JP
dc.textversion publisher ja_JP
dc.identifier.ncid AA0060007X ja_JP
dc.identifier.jtitle Cell Structure and Function ja_JP
dc.identifier.volume 43 ja_JP
dc.identifier.issue 2 ja_JP
dc.identifier.spage 153 ja_JP
dc.identifier.epage 169 ja_JP
dc.relation.doi info:doi/10.1247/csf.18012 ja_JP
dc.identifier.NAIST-ID 74654278 ja_JP
dc.relation.pmid 30047513 ja_JP

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