Imaging the early zebrafish embryo centrosomes following injection of small-molecule inhibitors to understand spindle formation.
Imaging the early zebrafish embryo centrosomes following injection of small-molecule inhibitors to understand spindle formation.
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DOI：
10.1016/j.xpro.2020.100293
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发表时间：
2021-03-19
期刊：
影响因子：
--
通讯作者：
Hehnly H
中科院分区：
文献类型：
--
作者：
Aljiboury AA;Mujcic A;Cammerino T;Rathbun LI;Hehnly H
在最早的分裂阶段，斑马鱼胚胎具有快速同步分裂的大细胞，在蛋黄顶部形成细胞层。在这里，我们描述了一个协议，用于监测纺锤体动力学在这些早期胚胎分裂。我们概述了注射斑马鱼胚胎与小分子抑制剂对polo样激酶，准备和安装胚胎的三维成像共聚焦显微镜技术。这些技术被用来了解早期斑马鱼胚胎的中心体如何构建有丝分裂纺锤体。有关本方案使用和执行的完整详细信息，请参阅。斑马鱼胚胎的培育和获取以监测纺锤体动力学斑马鱼胚胎显微注射以引入Polo样激酶（PLK 1/4）抑制剂安装活的和固定的斑马鱼胚胎用于早期胚胎研究使用共聚焦显微镜对活的和固定的斑马鱼胚胎进行成像在最早的分裂阶段，斑马鱼胚胎具有快速同步分裂的大细胞，在卵黄顶部产生细胞层。在这里，我们描述了一个协议，用于监测纺锤体动力学在这些早期胚胎分裂。我们概述了注射斑马鱼胚胎与小分子抑制剂对polo样激酶，准备和安装胚胎的三维成像共聚焦显微镜技术。这些技术被用来了解早期斑马鱼胚胎的中心体如何构建有丝分裂纺锤体。
During the earliest division stages, zebrafish embryos have large cells that divide rapidly and synchronously to create a cellular layer on top of the yolk. Here, we describe a protocol for monitoring spindle dynamics during these early embryonic divisions. We outline techniques for injecting zebrafish embryos with small-molecule inhibitors toward polo-like kinases, preparing and mounting embryos for three-dimensional imaging using confocal microscopy. These techniques are used to understand how the early zebrafish embryo’s centrosome constructs the mitotic spindle. For complete details on the use and execution of this protocol, please refer to. Breeding and obtaining zebrafish embryos to monitor spindle dynamics Zebrafish embryo microinjection to introduce polo-like kinase (PLK1/4) inhibitors Mounting of live and fixed zebrafish embryo for early embryonic studies Imaging live and fixed zebrafish embryos using confocal microscope During the earliest division stages, zebrafish embryos have large cells that divide rapidly and synchronously to create a cellular layer on top of the yolk. Here, we describe a protocol for monitoring spindle dynamics during these early embryonic divisions. We outline techniques for injecting zebrafish embryos with small-molecule inhibitors toward polo-like kinases, preparing and mounting embryos for three-dimensional imaging using confocal microscopy. These techniques are used to understand how the early zebrafish embryo’s centrosome constructs the mitotic spindle.
DOI：
10.1016/j.cub.2020.08.074
发表时间：
2020-11-16
期刊：
Current biology : CB
影响因子：
--
作者：
Rathbun LI;Aljiboury AA;Bai X;Hall NA;Manikas J;Amack JD;Bembenek JN;Hehnly H
通讯作者：
Hehnly H
影响因子：
2.5
作者：
KIMMEL, CB;BALLARD, WW;SCHILLING, TF
通讯作者：
SCHILLING, TF
影响因子：
48
作者：
Schindelin, Johannes;Arganda-Carreras, Ignacio;Frise, Erwin;Kaynig, Verena;Longair, Mark;Pietzsch, Tobias;Preibisch, Stephan;Rueden, Curtis;Saalfeld, Stephan;Schmid, Benjamin;Tinevez, Jean-Yves;White, Daniel James;Hartenstein, Volker;Eliceiri, Kevin;Tomancak, Pavel;Cardona, Albert
通讯作者：
Cardona, Albert