Automated analysis of time-lapse imaging of nuclear translocation by retrospective strategy and its application to STAT1 in HeLa cells.
Automated analysis of time-lapse imaging of nuclear translocation by retrospective strategy and its application to STAT1 in HeLa cells.
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回顾性策略对核转运的延时成像的自动分析及其在HELA细胞中的STAT1的应用。
DOI：
10.1371/journal.pone.0027454
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Zhang B
中科院分区：
文献类型：
--
作者：
Han F;Liang P;Wang F;Zeng L;Zhang B
基于细胞的延时成像图像分析主要受到荧光微弱和目标细胞结构边界模糊的挑战。为了解决这些瓶颈，研究人员开发了一种基于“回顾性”分析的新方法，用于在延时成像过程中发生微小形态变化的细胞。实验结束后用核染料对细胞进行固定和染色，对核染色图像进行分割得到的二值掩模对延时图像进行处理。这种自动化方法还可以识别在延时成像过程中移动的细胞，这是一个可能影响主要存在于细胞质中的靶蛋白的动力学测量的因素。然后，我们通过测量干扰素γ (IFNγ)诱导的信号转导器和活HeLa细胞中转录1激活因子（STAT1）核易位来验证该方法。我们的新方法首次实现了活细胞核易位的大规模自动化分析。细胞对IFNγ的反应在整个群体中表现出显著的漂移，但这些反应的共同特征使我们提出了STAT1输入的三阶段模型。该方法的简单性和自动化应使其能够应用于核细胞质易位的广谱延时研究。
Cell-based image analysis of time-lapse imaging is mainly challenged by faint fluorescence and dim boundaries of cellular structures of interest. To resolve these bottlenecks, a novel method was developed based on “retrospective” analysis for cells undergoing minor morphological changes during time-lapse imaging. We fixed and stained the cells with a nuclear dye at the end of the experiment, and processed the time-lapse images using the binary masks obtained by segmenting the nuclear-stained image. This automated method also identifies cells that move during the time-lapse imaging, which is a factor that could influence the kinetics measured for target proteins that are present mostly in the cytoplasm. We then validated the method by measuring interferon gamma (IFNγ) induced signal transducers and activators of transcription 1 (STAT1) nuclear translocation in living HeLa cells. For the first time, automated large-scale analysis of nuclear translocation in living cells was achieved by our novel method. The responses of the cells to IFNγ exhibited a significant drift across the population, but common features of the responses led us to propose a three-stage model of STAT1 import. The simplicity and automation of this method should enable its application in a broad spectrum of time-lapse studies of nuclear-cytoplasmic translocation.
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影响因子：
10.4
作者：
Kaltreider RC;Davis AM;Lariviere JP;Hamilton JW
通讯作者：
Hamilton JW
影响因子：
12.3
作者：
Carpenter AE;Jones TR;Lamprecht MR;Clarke C;Kang IH;Friman O;Guertin DA;Chang JH;Lindquist RA;Moffat J;Golland P;Sabatini DM
通讯作者：
Sabatini DM
DOI：
10.1080/03610927708827533
发表时间：
1977-01-01
期刊：
COMMUNICATIONS IN STATISTICS PART A-THEORY AND METHODS
影响因子：
--
作者：
HOLLAND, PW;WELSCH, RE
通讯作者：
WELSCH, RE
DOI：
10.1007/978-1-61779-243-4_18
发表时间：
2011-01-01
期刊：
ANDROGEN ACTION: METHODS AND PROTOCOLS
影响因子：
--
作者：
Hartig, Sean M.;Newberg, Justin Y.;Mancini, Michael A.
通讯作者：
Mancini, Michael A.
影响因子：
7.3
作者：
Kodiha, Mohamed;Brown, Claire M.;Stochaj, Ursula
通讯作者：
Stochaj, Ursula