人外周血CD4^+T细胞F-actin的超分辨SIM成像

Super-resolution structured illumination microscopy(SIM)imaging of F-actin in human peripheral blood CD4^+T cells

通过合成lifeact序列,将其克隆到pEGFP-C1载体上,构建了pEGFP-C1-Lifeact质粒,转染原代CD4+T细胞,实现对CD4+T细胞F-actin的荧光标记,结合超分辨3D-SIM荧光成像技术,研究人外周血CD4+T细胞F-actin的精细结构,并对其动力学过程进行连续观察。结果表明,CD4+T细胞F-actin分布在细胞膜周围,处于解聚和聚合的动态平衡状态,与普通荧光显微镜相比,超分辩SIM成像F-actin结构更加清晰,分辨率提高2~4倍。

To label the CD4^+T cells F-actin,the lifeact gene was synthesized and cloned into vector pEGFP-C1,CD4^+T cells were then transfected with pEGFP-C1-Lifeact plasmid and imaged by the real time super-resolution 3D-SIM fluorescence microscopy.The results showed that F-actin is distributed around the cellular membrane with steady state of depolymerization and polymerization.It was indicated that the spatial resolution of the fine SIM imaging of F-actin was two to four times higher than the wide field fluorescence microscope imaging.