Automated analysis of invadopodia dynamics in live cells.
Automated analysis of invadopodia dynamics in live cells.
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作者：
Berginski ME;Creed SJ;Cochran S;Roadcap DW;Bear JE;Gomez SM
多种细胞类型形成专门的蛋白质复合体，细胞利用这些复合体主动降解周围的细胞外基质。这些结构被称为足体或内足体，统称为侵入体。由于它们在健康生理学以及癌症等病理条件下的潜在重要性，这些结构的特征越来越引起人们的兴趣。根据早期对内翻足的描述，开发了标记转移癌细胞下的基质的分析方法，允许评估内翻足在运动细胞中的活性。然而，使用这些方法对失足动物的表征传统上是用耗时且可能存在偏见的量化方法手动完成的，这限制了实验的数量和可以分析的数据量。我们已经开发了一个系统来自动分割、跟踪和量化单个和整个细胞水平上的延时荧光图像集中的隐足动物。我们严格测试了该方法通过使用具有良好特征的小分子抑制剂来检测内侧足的形成和动态变化的能力，这些小分子抑制剂对内足的影响是已知的。我们的结果证明了这种分析方法能够以高通量、自动化的方式从活细胞成像数据中量化失足形成的变化。
Multiple cell types form specialized protein complexes that are used by the cell to actively degrade the surrounding extracellular matrix. These structures are called podosomes or invadopodia and collectively referred to as invadosomes. Due to their potential importance in both healthy physiology as well as in pathological conditions such as cancer, the characterization of these structures has been of increasing interest. Following early descriptions of invadopodia, assays were developed which labelled the matrix underneath metastatic cancer cells allowing for the assessment of invadopodia activity in motile cells. However, characterization of invadopodia using these methods has traditionally been done manually with time-consuming and potentially biased quantification methods, limiting the number of experiments and the quantity of data that can be analysed. We have developed a system to automate the segmentation, tracking and quantification of invadopodia in time-lapse fluorescence image sets at both the single invadopodia level and whole cell level. We rigorously tested the ability of the method to detect changes in invadopodia formation and dynamics through the use of well-characterized small molecule inhibitors, with known effects on invadopodia. Our results demonstrate the ability of this analysis method to quantify changes in invadopodia formation from live cell imaging data in a high throughput, automated manner.
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DOI：
10.1038/nrm3141
发表时间：
2011-06-23
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
通讯作者：
--
影响因子：
9.2
作者：
Sharma, Ved P.;Eddy, Robert;Entenberg, David;Kai, Masayuki;Gertler, Frank B.;Condeelis, John
通讯作者：
Condeelis, John
影响因子：
3.3
作者：
Beaty BT;Sharma VP;Bravo-Cordero JJ;Simpson MA;Eddy RJ;Koleske AJ;Condeelis J
通讯作者：
Condeelis J
影响因子：
48
作者：
Riedl, Julia;Crevenna, Alvaro H.;Wedlich-Soldner, Roland
通讯作者：
Wedlich-Soldner, Roland
影响因子：
9.2
作者：
Tang, Haoran;Li, Ang;Machesky, Laura M.
通讯作者：
Machesky, Laura M.