Image analysis for automatic segmentation of cytoplasms and classification of Rac1 activation
Image analysis for automatic segmentation of cytoplasms and classification of Rac1 activation
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DOI：
10.1002/cyto.a.10107
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发表时间：
2004-01-01
期刊：
影响因子：
3.7
通讯作者：
Zaltsman, A
中科院分区：
文献类型：
--
作者：
Lindblad, J;W채hlby, C;Zaltsman, A
背景：Rac1是一种gtp结合分子，参与广泛的细胞过程。利用数字图像分析，可以定量估计激动剂诱导的绿色荧光蛋白（GFP） Rac1向细胞膜的易位。方法：描述了一种全自动图像分析方法，根据刺激下GFP-Rac1易位和皱褶形成的激活情况对细胞进行分割、特征提取和分类。基于四幅图像序列视觉标注产生的训练数据，建立统计分类器。结果：将自动分类结果与同一时间序列目测结果进行了比较。自动分类与视觉分类在同一水平上存在差异，就像两种不同技术人员的视觉分类存在差异一样。对含有7幅不同激动剂浓度的图像序列的第二组图像集进行分类，结果表明，当激动剂浓度增加时，分类器可以检测到激活细胞的比例增加。结论：细胞内活动，如皱褶的形成，可以通过全自动图像分析来量化，其准确性与目测相当。这种分析可以以每秒数百个细胞的速度完成，并且没有人工判断引入的主观性。(C) 2003 Wiley-Liss, Inc。
Background: Rac1 is a GTP-binding molecule involved in a wide range of cellular processes. Using digital image analysis, agonist-induced translocation of green fluorescent protein (GFP) Rac1 to the cellular membrane can be estimated quantitatively for individual cells.Methods: A fully automatic image analysis method for cell segmentation, feature extraction, and classification of cells according to their activation, i.e., GFP-Rac1 translocation and ruffle formation at stimuli, is described. Based on training data produced by visual annotation of four image series, a statistical classifier was created.Results: The results of the automatic classification were compared with results from visual inspection of the same time sequences. The automatic classification differed from the visual classification at about the same level as visual classifications performed by two different skilled professionals differed from each other. Classification of a second image set, consisting of seven image series with different concentrations of agonist, showed that the classifier could detect an increased proportion of activated cells at increased agonist concentration.Conclusions: Intracellular activities, such as ruffle formation, can be quantified by fully automatic image analysis, with an accuracy comparable to that achieved by visual inspection. This analysis can be done at a speed of hundreds of cells per second and without the subjectivity introduced by manual judgments. (C) 2003 Wiley-Liss, Inc.