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DOI: 10.3791/52343-v
Please note that some of the translations on this page are AI generated. Click here for the English version.
在肿瘤微机械刚性起着通过增加侵袭伪足活性和肌动球蛋白收缩驱动恶性行为了至关重要的作用。用聚丙烯酰胺凝胶(部分分配),侵袭伪足和牵引力测定可以被用于研究癌细胞的侵入性和收缩特性响应于矩阵刚性。
该程序的总体目标是创建可用于入侵者、足和牵引力测定的聚丙烯酰胺凝胶,以研究癌细胞的侵袭性和收缩性。这是通过首先激活玻璃花盘中的 14 毫米盖玻片来实现的,以确保聚丙烯酰胺凝胶与玻璃结合。接下来,制备用于两种分析的聚丙烯酰胺溶液,以产生具有不同刚性或机械性能的凝胶。
然后用一层薄薄的荧光标记的细胞外基质制备侵袭的 podia 测定以检测降解。最后,用癌细胞接种测定进行过夜孵育,然后固定和染色入侵者、足板和牵引力板的活细胞成像。最终,荧光显微镜用于对入侵者、足和细胞外基质降解进行成像。
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