Microtubule organization within mitotic spindles revealed by serial block face scanning electron microscopy and image analysis.

Microtubule organization within mitotic spindles revealed by serial block face scanning electron microscopy and image analysis.
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DOI:
10.1242/jcs.203877
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发表时间:
2017-05-15
影响因子:
4
通讯作者:
Royle SJ
Royle SJ
中科院分区:
生物学2区
文献类型:
--
作者:
Nixon FM;Honnor TR;Clarke NI;Starling GP;Beckett AJ;Johansen AM;Brettschneider JA;Prior IA;Royle SJ

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连续块面扫描电子显微镜(SBF-SEM)是一种在3D中分析细胞的强大方法。在这里,在该方法的分辨率极限工作,我们描述了一个相关的光-SBF-SEM工作流程,以解决人类细胞中的有丝分裂纺锤体微管。我们提出了四个例子,使用这个工作流程是不实际的光学显微镜和/或透射电子显微镜。首先,区分K-纤维内密切相关的微管;第二,解决有丝分裂纺锤体中的桥接纤维;第三,可视化有丝分裂细胞中的膜,相对于纺锤体装置;第四,着丝粒的体积分析。我们的工作流程还包括新的计算工具,用于探索有丝分裂纺锤体内微管的空间排列。我们使用这些工具来显示有丝分裂纺锤体中微管的顺序对纺锤体上TACC 3的水平敏感。
Serial block face scanning electron microscopy (SBF-SEM) is a powerful method to analyze cells in 3D. Here, working at the resolution limit of the method, we describe a correlative light–SBF-SEM workflow to resolve microtubules of the mitotic spindle in human cells. We present four examples of uses for this workflow that are not practical by light microscopy and/or transmission electron microscopy. First, distinguishing closely associated microtubules within K-fibers; second, resolving bridging fibers in the mitotic spindle; third, visualizing membranes in mitotic cells, relative to the spindle apparatus; and fourth, volumetric analysis of kinetochores. Our workflow also includes new computational tools for exploring the spatial arrangement of microtubules within the mitotic spindle. We use these tools to show that microtubule order in mitotic spindles is sensitive to the level of TACC3 on the spindle.
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