A Quantitative Image Cytometry Technique for Time Series or Population Analyses of Signaling Networks 英文参考文献.docVIP

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A Quantitative Image Cytometry Technique for Time Series or Population Analyses of Signaling Networks 英文参考文献.doc

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A Quantitative Image Cytometry Technique for Time Series or Population Analyses of Signaling Networks 英文参考文献

AQuantitativeImageCytometryTechniqueforTime SeriesorPopulationAnalysesofSignalingNetworks Yu-ichiOzaki1*,ShinsukeUda1,TakeshiH.Saito1,JaehoonChung1,HiroyukiKubota1,ShinyaKuroda1,2* 1DepartmentofBiophysicsandBiochemistry,GraduateSchoolofScience,UniversityofTokyo,Bunkyo-ku,Tokyo,Japan,2CREST,JapanScienceandTechnologyAgency, UniversityofTokyo,Bunkyo-ku,Tokyo,Japan Abstract Background:Modelingofcellularfunctionsonthebasisofexperimentalobservationisincreasinglycommoninthefieldof cellular signaling. However, such modeling requires a large amount of quantitative data of signaling events with high spatio-temporalresolution.Anoveltechniquewhichallowsustoobtainsuchdataisneededforsystemsbiologyofcellular signaling. Methodology/Principal Findings: We developed a fully automatable assay technique, termed quantitative image cytometry (QIC), which integrates a quantitative immunostaining technique and a high precision image-processing algorithmforcellidentification.Withtheaidofanautomatedsamplepreparationsystem,thisdevicecanquantifyprotein expression, phosphorylation and localization with subcellular resolution at one-minute intervals. The signaling activities quantified by the assay system showed good correlation with, as well as comparable reproducibility to, western blot analysis. Taking advantage of the high spatio-temporal resolution, we investigated the signaling dynamics of the ERK pathwayinPC12cells. Conclusions/Significance: The QIC technique appears as a highly quantitative and versatile technique, which can be a convenientreplacementforthemostconventionaltechniquesincludingwesternblot,flowcytometryandlivecellimaging. Thus,theQICtechniquecanbeapowerfultoolforinvestigatingthesystemsbiologyofcellularsignaling. Citation: Ozaki Y-i, Uda S, Saito TH, Chung J, Kubota H, et al. (2010) A Quantitative Image Cytometry Technique for Time Series or Population Analyses of SignalingNetworks.PLoSONE5(4):e9955.doi:10.1371/journal.pone.0009955 Editor:JeanPeccoud,VirginiaTech,UnitedSta