a quantitative image cytometry technique for time series or population analyses of signaling networks定量图像时间序列的血细胞计数技术或信号网络的人口分析.pdfVIP

A Quantitative Image Cytometry Technique for Time Series or Population Analyses of Signaling Networks 1 1 1 1 1 1,2 Yu-ichi Ozaki *, Shinsuke Uda , Takeshi H. Saito , Jaehoon Chung , Hiroyuki Kubota , Shinya Kuroda * 1 Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo, Japan, 2 CREST, Japan Science and Technology Agency, University of Tokyo, Bunkyo-ku, Tokyo, Japan Abstract Background: Modeling of cellular functions on the basis of experimental observation is increasingly common in the field of cellular signaling. However, such modeling requires a large amount of quantitative data of signaling events with high spatio-temporal resolution. A novel technique which allows us to obtain such data is needed for systems biology of cellular 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 algorithm for cell identification. With the aid of an automated sample preparation system, this device can quantify protein 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 pathway in PC12 cells. Conclusions/S