A versatile multivariate image analysis pipeline reveals features of Xenopus extract spindles.
A versatile multivariate image analysis pipeline reveals features of Xenopus extract spindles.
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DOI：
10.1083/jcb.201509079
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发表时间：
2016-04-11
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影响因子：
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通讯作者：
Heald R
中科院分区：
文献类型：
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作者：
Grenfell AW;Strzelecka M;Crowder ME;Helmke KJ;Schlaitz AL;Heald R
作者描述了自动图像和数据分析工具，揭示了非洲爪蟾卵提取物纺锤体的结构原理，允许对纺锤体表型进行快速，无偏见的评估，并可用于分析其他亚细胞结构，如细胞核。成像数据集具有丰富的定量信息。然而，很少有细胞生物学家拥有必要的工具来分析它们。在这里，我们提出了一个大型数据集的爪蟾提取物纺锤体图像与分析管道，旨在评估纺锤体形态在一系列实验条件。我们对不同纺锤体类型的分析说明了动粒微管是如何放大纺锤体微管密度的。提取物混合实验表明，一些纺锤体功能滴定，而其他经历开关样的过渡，和多变量分析显示的多效性形态的影响，调制TPX2的水平，一个关键的纺锤体组装因子。我们还应用我们的管道来分析人类细胞培养中的核形态，显示分割方法的一般实用性。我们的分析为纺锤体类型的多样性提供了新的见解，并为未来的研究提出了建议。所概述的方法可以应用于其他研究人员研究纺锤体形态和适应与其他实验系统的最小修改。
The authors describe automated image and data analysis tools that reveal architectural principles of the Xenopus egg extract spindle, allow for rapid, unbiased assessment of spindle phenotypes, and can be adapted to analyze other subcellular structures such as nuclei. Imaging datasets are rich in quantitative information. However, few cell biologists possess the tools necessary to analyze them. Here, we present a large dataset of Xenopus extract spindle images together with an analysis pipeline designed to assess spindle morphology across a range of experimental conditions. Our analysis of different spindle types illustrates how kinetochore microtubules amplify spindle microtubule density. Extract mixing experiments reveal that some spindle features titrate, while others undergo switch-like transitions, and multivariate analysis shows the pleiotropic morphological effects of modulating the levels of TPX2, a key spindle assembly factor. We also apply our pipeline to analyze nuclear morphology in human cell culture, showing the general utility of the segmentation approach. Our analyses provide new insight into the diversity of spindle types and suggest areas for future study. The approaches outlined can be applied by other researchers studying spindle morphology and adapted with minimal modification to other experimental systems.