Cell Lineage Reconstruction of Early Zebrafish Embryos Using Label-Free Nonlinear Microscopy
Cell Lineage Reconstruction of Early Zebrafish Embryos Using Label-Free Nonlinear Microscopy
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DOI：
10.1126/science.1189428
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发表时间：
2010-08-20
期刊：
影响因子：
56.9
通讯作者：
Beaurepaire, Emmanuel
中科院分区：
文献类型：
--
作者：
Olivier, Nicolas;Luengo-Oroz, Miguel A.;Beaurepaire, Emmanuel
量化动物早期胚胎发生中的细胞行为仍然是一个具有挑战性的问题，需要整体成像和自动图像分析。我们设计了一个框架，用于对未染色的整个斑马鱼胚胎进行前 10 个细胞分裂周期的成像和重建，并以微米级空间分辨率和分钟级时间精度报告沿细胞谱系的测量结果。点扫描多光子激发经过优化，可优先探测胚胎的最内部区域，提供突出显示所有有丝分裂纺锤体和细胞边界的内在信号。自动图像分析揭示了细胞增殖的现象。卵裂球不断地不同步。 32细胞阶段后，细胞周期根据细胞径向位置延长，导致明显的分裂波。与系统动力学突变的经典描述相比，这个过程的渐进放大是规则。
Quantifying cell behaviors in animal early embryogenesis remains a challenging issue requiring in toto imaging and automated image analysis. We designed a framework for imaging and reconstructing unstained whole zebrafish embryos for their first 10 cell division cycles and report measurements along the cell lineage with micrometer spatial resolution and minute temporal accuracy. Point-scanning multiphoton excitation optimized to preferentially probe the innermost regions of the embryo provided intrinsic signals highlighting all mitotic spindles and cell boundaries. Automated image analysis revealed the phenomenology of cell proliferation. Blastomeres continuously drift out of synchrony. After the 32-cell stage, the cell cycle lengthens according to cell radial position, leading to apparent division waves. Progressive amplification of this process is the rule, contrasting with classical descriptions of abrupt changes in the system dynamics.