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Author:
Date:
2016-01-05
[Abstract] Cell migration is a highly complex and dynamic biological process, essential in several physiological phenomena and pathologies including cancer dissemination and metastasis formation. Thus understanding single cell migration is highly relevant and requires a suitable image-based assay. Depending on the speed of the moving cells, one may require fast time-lapse microscopy, which is not always suitable for high-throughput screening. To overcome this, a quantitative and fixed single cell migration assay was developed based on the PhagoKinetic Tracks (PKT) procedure. Briefly, cells are seeded on top of a monolayer of carboxylated latex beads, and as cells migrate, they phagocytose these beads and leave behind a migratory track. These bead-free migratory tracks can be visualized using a ...
[摘要] 细胞迁移是高度复杂和动态的生物过程，在几种生理现象和病理包括癌症传播和转移形成中是必需的。因此，理解单细胞迁移是高度相关的，并需要合适的基于图像的测定。根据移动细胞的速度，可能需要快速时间延迟显微术，这不总是适合于高通量筛选。为了克服这一点，基于Phago运动轨迹（PKT）程序开发了定量和固定的单细胞迁移测定。简言之，将细胞接种在单层羧化的乳胶珠的顶部，并且当细胞迁移时，它们吞噬这些珠并留下迁移轨迹。这些无珠移动轨迹可以使用标准明场显微镜可视化，并分析单细胞迁移的多参数定量评估（Naffar-Abu-Amara等人，2008）。
在这里，我们描述了PKT测定的详细和优化的方案，适用于RNAi和药物筛选（van Roosmalen等人，2015）。这个协议允许用户在单细胞水平研究迁移行为，没有快速和活成像显微镜。
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