小鼠胚胎的4D光片成像、计算重建和细胞跟踪。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-01-02 DOI:10.1016/j.xpro.2024.103515

Martin H Dominguez, Jonathon M Muncie-Vasic, Benoit G Bruneau

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引用次数: 0

摘要

随着薄片荧光显微镜（LSFM）的广泛应用，延时成像的重建将进一步加深我们对细胞分辨率下复杂生物过程的理解。在这里，我们提出了一个全面的工作流程，用于捕获，处理和分析多视图LSFM实验，使用离体小鼠胚胎作为发育模型系统。我们的方案描述了在商用LSFM仪器上的成像，然后使用开源软件对离散段进行计算分析。定量迁移和形态动力学包括在内。有关使用和执行该协议的完整细节，请参阅Dominguez et al.1。

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4D light sheet imaging, computational reconstruction, and cell tracking in mouse embryos.

As light sheet fluorescence microscopy (LSFM) becomes widely available, reconstruction of time-lapse imaging will further our understanding of complex biological processes at cellular resolution. Here, we present a comprehensive workflow for in toto capture, processing, and analysis of multi-view LSFM experiments using the ex vivo mouse embryo as a model system of development. Our protocol describes imaging on a commercial LSFM instrument followed by computational analysis in discrete segments, using open-source software. Quantification of migration and morphodynamics is included. For complete details on the use and execution of this protocol, please refer to Dominguez et al.¹.

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