Using total internal reflection fluorescence (TIRF) microscopy to visualize cortical actin and microtubules in the Drosophila syncytial embryo.

Using total internal reflection fluorescence (TIRF) microscopy to visualize cortical actin and microtubules in the Drosophila syncytial embryo.
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DOI:
10.1002/dvdy.22076
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发表时间:
2009-10
影响因子:
2.5
通讯作者:
McCartney, Brooke M.
McCartney, Brooke M.
中科院分区:
生物学3区
文献类型:
--
作者:
Webb, Rebecca L.;Rozov, Orr;Watkins, Simon C.;McCartney, Brooke M.

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果蝇合胞体胚胎是研究动态协调细胞骨架重排的一个强有力的发育模型系统。共聚焦显微镜已经开始揭示更多关于胚胎发生过程中细胞骨架的变化。全内反射荧光(TIRF)显微镜提供了一个很有前途的新方法,提高轴向分辨率的可视化皮质事件。我们已经应用TIRF显微镜观察果蝇胚胎皮层微管和肌动蛋白在合胞胚盘的动态。在这里,我们描述了这种技术的细节,并报告皮质微管和肌动蛋白在果蝇合胞体胚胎的定性评估。此外,我们确定了一个高峰期的皮质微管在后期的每个核周期的合胞胚盘,并使用TIRF显微镜产生的图像,我们定量分析微管动力学在这段时间。
The Drosophila syncytial embryo is a powerful developmental model system for studying dynamic coordinated cytoskeletal rearrangements. Confocal microscopy has begun to reveal more about the cytoskeletal changes that occur during embryogenesis. Total Internal Reflection Fluorescence (TIRF) microscopy provides a promising new approach for the visualization of cortical events with heightened axial resolution. We have applied TIRF microscopy to the Drosophila embryo to visualize cortical microtubule and actin dynamics in the syncytial blastoderm. Here we describe the details of this technique, and report qualitative assessments of cortical microtubules and actin in the Drosophila syncytial embryo. In addition, we identified a peak of cortical microtubules during anaphase of each nuclear cycle in the syncytial blastoderm, and using images generated by TIRF microscopy we quantitatively analyzed microtubule dynamics during this time.
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