声穿孔
钙黄绿素
生物物理学
微气泡
细胞膜
细胞内
HEK 293细胞
细胞质
化学
流出
细胞
荧光显微镜
单细胞分析
膜
细胞生物学
荧光
超声波
生物化学
生物
物理
量子力学
基因
声学
作者
Zhenzhen Fan,Haiyan Liu,Michael Mayer,Cheri X. Deng
标识
DOI:10.1073/pnas.1208198109
摘要
This paper presents unique approaches to enable control and quantification of ultrasound-mediated cell membrane disruption, or sonoporation, at the single-cell level. Ultrasound excitation of microbubbles that were targeted to the plasma membrane of HEK-293 cells generated spatially and temporally controlled membrane disruption with high repeatability. Using whole-cell patch clamp recording combined with fluorescence microscopy, we obtained time-resolved measurements of single-cell sonoporation and quantified the size and resealing rate of pores. We measured the intracellular diffusion coefficient of cytoplasmic RNA/DNA from sonoporation-induced transport of an intercalating fluorescent dye into and within single cells. We achieved spatiotemporally controlled delivery with subcellular precision and calcium signaling in targeted cells by selective excitation of microbubbles. Finally, we utilized sonoporation to deliver calcein, a membrane-impermeant substrate of multidrug resistance protein-1 (MRP1), into HEK-MRP1 cells, which overexpress MRP1, and monitored the calcein efflux by MRP1. This approach made it possible to measure the efflux rate in individual cells and to compare it directly to the efflux rate in parental control cells that do not express MRP1.
科研通智能强力驱动
Strongly Powered by AbleSci AI