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6.7K Views
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11:07 min
August 14th, 2019
DOI :
10.3791/59849-v
Chapters
0:06
Title
0:44
Preparation of Embryos for Implantation
1:31
Perivitelline Injection of Mammalian Cancer Cells into 2 dpf (Day Post Fertilization) Embryos
3:55
Live Imaging and Quantitation of the Angiogenic Response to the Zebrafish Xenograft
8:34
Results: Quantitation of the Host Angiogenic Response in Zebrafish Tumor Xenografts
10:37
Conclusion
Transcript
该协议描述了转基因斑马鱼幼虫如何可用于提供肿瘤异种血管化的体内定量测定。与以前的技术相比,该技术的主要优点是,它允许研究者准确量化异种血管化水平的变化。关键步骤是确保肿瘤细胞正确注射到斑马鱼幼虫中,以便随后的成像和定量步骤不会受到影响。
在生长和标记B16-F1细胞与荧光染料后,开始准备胚胎植入。在E3溶液中用2%的甲基纤维素填充35毫米的盘子,达到盘子体积的四分之一。使用转移移液器,将大约 50 个以前准备的转基因胚胎放在甲基纤维素上,同时最大限度地减少 E3 溶液的转移量。
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Summary
这种方法的目的是通过异种移植哺乳动物肿瘤细胞进入带有荧光标记血管的斑马鱼胚胎,生成肿瘤血管生成体内模型。通过成像异种移植物和相关血管,可以得到血管原反应的定量测量。
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