An ensemble-averaged, cell density-based digital model of zebrafish embryo development derived from light-sheet microscopy data with single-cell resolution.
An ensemble-averaged, cell density-based digital model of zebrafish embryo development derived from light-sheet microscopy data with single-cell resolution.
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DOI：
10.1038/srep08601
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发表时间：
2015-02-25
影响因子：
4.6
通讯作者：
Nienhaus GU
中科院分区：
文献类型：
--
作者：
Kobitski AY;Otte JC;Takamiya M;Schäfer B;Mertes J;Stegmaier J;Rastegar S;Rindone F;Hartmann V;Stotzka R;García A;van Wezel J;Mikut R;Strähle U;Nienhaus GU
生物全细胞形态发生的定量成像技术的最新进展开启了发育生物学的新纪元。在这里，我们开发了一种基于单细胞分辨率的光片荧光显微镜框架，用于识别和表征毫米级生物的细微表型变化。这样的比较研究需要对整个总体的分析，以便能够区分样本之间的差异和明确的表型变化。我们提出了一个斑马鱼胚胎发育到16 h的动力学数字模型。该模型基于对多个个体采集的数据的精确叠加和平均，并描述了每个时间点的细胞密度及其迁移方向。多样本比较研究的定量指标已被引入，以分析群体内的发育差异。数字模型可以作为一张画布，在上面可以研究细胞亚群的行为。作为一个例子，我们研究了原肠形成开始时胚层形成过程中的细胞重排。将独眼针头突变体(OEP)与野生型胚胎的数字模型进行比较，发现它在原肠形成开始时发育异常，几个小时后眼睛才能看到明显的变化。
A new era in developmental biology has been ushered in by recent advances in the quantitative imaging of all-cell morphogenesis in living organisms. Here we have developed a light-sheet fluorescence microscopy-based framework with single-cell resolution for identification and characterization of subtle phenotypical changes of millimeter-sized organisms. Such a comparative study requires analyses of entire ensembles to be able to distinguish sample-to-sample variations from definitive phenotypical changes. We present a kinetic digital model of zebrafish embryos up to 16 h of development. The model is based on the precise overlay and averaging of data taken on multiple individuals and describes the cell density and its migration direction at every point in time. Quantitative metrics for multi-sample comparative studies have been introduced to analyze developmental variations within the ensemble. The digital model may serve as a canvas on which the behavior of cellular subpopulations can be studied. As an example, we have investigated cellular rearrangements during germ layer formation at the onset of gastrulation. A comparison of the one-eyed pinhead (oep) mutant with the digital model of the wild-type embryo reveals its abnormal development at the onset of gastrulation, many hours before changes are obvious to the eye.