原子力声显微镜评价超声靶向微泡破坏法对大鼠心肌母细胞的影响  

Assessment of Effect of Ultrasound Targeted Microbubble Destruction on Rat Myocardial Cells by Atomic Force Acoustic Microcope

作　　者：金佳美[1] 张宇辉[1] 陈明[1] 钱梦騄[2] 程茜[2] 陈红[2] 

机构地区：[1]同济大学附属东方医院心超室,上海市200120 [2]同济大学声学研究所

出　　处：《中国超声医学杂志》2013年第10期929-933,共5页Chinese Journal of Ultrasound in Medicine

基　　金：国家自然科学基金青年科学基金项目(No.10804085;10774113);上海市卫生局青年基金项目(No.2010Y012);上海市浦东新区卫生系统优秀青年医学人才项目(No.PW Rq201004)

摘　　要：目的探讨运用原子力声显微镜评价超声靶向微泡破坏法对大鼠心肌母细胞(H9C2)影响的可行性。方法采用不同超声参数对H9C2细胞进行辐照处理。使用原子力声显微镜观察细胞骨架、形态结构的改变;采用台盼蓝染色法及荧光物质转入法检测细胞存活率及细胞膜通透性变化。结果(1)通过比较辐照后H9C2细胞生存率及发光率变化,适宜参数为:频率1 MHz,声强1.0 W/cm^2,占空比20%,辐照时间60 s;(2)原子力声显微镜可以完整显示H9C2细胞的形态结构,以形态像、声学像及三维像的差异区分细胞损伤的程度。结论原子力声显微镜能够准确评估不同超声辐照参数对H9C2细胞的影响,优化超声靶向微泡破坏法转染参数。Objective To assess effect of ultrasound targeted microbubble destruction（UTMD） on H9C2 cells by atomic force acoustic microeope（AFAM）. Methods H9C2 cells were exposed to ultrasound radiation with different parameters. Then cells were imaged by AFAM for observing the change of cell structure and cytoskeieton. And cells vi- ability and membrane permeability were tested by trypan blue exclusion and fluorescent substance. Results （1） The optimal ultrasound parameters ： frequency 1 MHz, intensity 1.0 W/cm2 , Duty cycle 20%, irradiation time 60 s. The conclusion was drawn from comparison of cell viability and luminosity factor of cells treated with different ultrasound parameters. （2）Cytomembrane, cytoplasm, eytoskeleton and cell nucleus in H9C2 cells could be imaged clearly and completely by AFAM. Different degree of cell injury could be distinguished by form view, acoustic view and three-dimensional view of AFAM. Conclusions The effect of UTMD combined with different ultrasound parameters on cell structure and cytoskeleton of H9C2 cells could be evaluated by AFAM effectively. And the transfection parameters of UTMD could be optimized by AFAM assessment.

关 键 词：原子力声显微镜 超声靶向微泡破坏法 细胞骨架 大鼠心肌母细胞 

分 类 号：R445.1[医药卫生—影像医学与核医学]