A Quantitative Image Cytometry Technique for Time Series or Population Analyses of Signaling Networks
A Quantitative Image Cytometry Technique for Time Series or Population Analyses of Signaling Networks
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DOI：
10.1371/journal.pone.0009955
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发表时间：
2010-04-01
期刊：
影响因子：
3.7
通讯作者：
Kuroda, Shinya
中科院分区：
文献类型：
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作者：
Ozaki, Yu-ichi;Uda, Shinsuke;Kuroda, Shinya
背景：基于实验观察的细胞功能建模在细胞信号传导领域越来越普遍。然而，这样的建模需要大量的具有高时空分辨率的信号事件的定量数据。一种新的技术，使我们能够获得这样的数据是需要系统生物学的cellularsignaling.Methodology/主要发现：我们开发了一种完全自动化的检测技术，称为定量图像细胞仪（QIC），它集成了定量免疫染色技术和高精度的图像处理算法的细胞identification。借助自动化样品制备系统，该设备可以以1分钟的间隔定量蛋白质表达、磷酸化和亚细胞分辨率定位。通过测定系统定量的信号传导活性显示出与蛋白质印迹分析的良好相关性以及可比较的再现性。利用高时空分辨率的优势，我们研究了ERK通路在PC 12 cells.Conclusions/Significance的信号动力学：QIC技术出现作为一个高度定量和通用的技术，它可以是一个方便的替代最传统的技术，包括Western blot，流式细胞术和活细胞成像。因此，QIC技术可以成为研究细胞信号传导的系统生物学的有力工具。
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/Significance: The QIC technique appears as a highly quantitative and versatile technique, which can be a convenient replacement for the most conventional techniques including western blot, flow cytometry and live cell imaging. Thus, the QIC technique can be a powerful tool for investigating the systems biology of cellular signaling.