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CO2 incubator
Company: BINDER
Catalog#: CB160
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Primed Track: Reliable Volumetric Single-cell Tracking and Lineage Tracing of Living Specimen with Dual-labeling Approaches
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2020-06-05
[Abstract]  Mammalian embryonic development starts with a single fertilized zygote that develops into a blastocyst embryo consisting of three cell types that evolve into either embryonic or extra-embryonic tissues. Lineage tracing of these cells can provide important information about the molecular and cellular dynamics contributing to fate allocation during early development. While global labeling techniques allow for visualization of all cells at the same time, lineage tracing of cells over several divisions can become complicated due to embryo movement and rotation as well as increasing cell densities. Here, we use green-to-red photoconvertible proteins for both global and sparse labeling of cells of interest in the developing murine embryo. We use primed conversion to achieve precise ... [摘要]  [摘要 ] 哺乳动物的胚胎发育始于单个受精的受精卵，受精卵发育为囊胚胚胎，由三种类型的细胞演化为胚胎或胚外组织，这些细胞的谱系追踪可提供有关有助于命运分配的分子和细胞动力学的重要信息。在早期开发过程中，虽然全局标记技术可以同时可视化所有细胞，但由于胚胎的移动和旋转以及细胞密度的增加，跨多个分区的细胞谱系追踪可能会变得复杂。红色光转换蛋白，用于在发育中的鼠胚胎中对目标细胞进行全局标记和稀疏标记。我们使用引物转换来实现4细胞期胚胎中单核的精确光转换，然后进行体积实时成像以捕获直至胚泡阶段的发育。我们开发了一个图像分析管道，称为 启动的Track ，使用双重标记策略对胚胎中的所有细胞进行直接的分段和配准，以及旋转和空间漂移的校正，共同实现了单个细胞的可靠，快速的跟踪和谱系追踪，甚至可以增加的成像时间间隔会导致数据量大幅减少，这是体积长期成像技术的所有必要条件。

[背景 ] 哺乳动物发育的第一谱系的最初几天内受精后注入时的胚胎附着于子宫壁之前指定（威灵等人，2016）.Still ，如何和何时这些第一专门的细胞类型出现，并且分子机制是造成最初均质细胞分裂的原因，目前尚不清楚。选择性平面照明显微镜（SPIM）提供了一种独特的手段，能够以无创方式连续几天捕获活样品的发育进程（Strnad 等等人，2016年 ; McDole ...

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