物理化学学报 >> 2012, Vol. 28 >> Issue (06): 1502-1508.doi: 10.3866/PKU.WHXB201203201

生物物理化学 上一篇    下一篇

基于AFM的药物刺激前后淋巴瘤活细胞的形貌及弹性的变化

李密1,2, 刘连庆1, 席宁1,3, 王越超1, 董再励1, 肖秀斌4, 张伟京4   

  1. 1. 中国科学院沈阳自动化研究所, 机器人学国家重点实验室, 沈阳 110016;
    2. 中国科学院研究生院, 北京 100049;
    3. 密歇根州立大学电子与计算机工程系, 东兰辛 48824, 美国;
    4. 军事医学科学院附属医院淋巴科, 北京 100071
  • 收稿日期:2012-03-09 修回日期:2012-03-20 发布日期:2012-05-17
  • 通讯作者: 刘连庆, 席宁, 张伟京 E-mail:lqliu@sia.cn; xin@egr.msu.edu; zhangwj3072@163.com
  • 基金资助:

    国家自然科学基金(60904095, 61175103), 国家高技术研究发展计划项目(863) (2009AA03Z316)及中国科学院、国家外国专家局创新团队国际合作伙伴计划资助

Drug-Induced Changes of Topography and Elasticity in Living B Lymphoma Cells Based on Atomic Force Microscopy

LI Mi1,2, LIU Lian-Qing1, XI Ning1,3, WANG Yue-Chao1, DONG Zai-Li1, XIAO Xiu-Bin4, ZHANG Wei-Jing4   

  1. 1. State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang 110016, P. R. China;
    2. Graduate University of Chinese Academy of Sciences, Beijing 100049, P. R. China;
    3. Department of Electrical and Computer Engineering, Michigan State University, East Lansing 48824, USA;
    4. Department of Lymphoma, Affiliated Hospital of Military Medical Academy of Sciences, Beijing 100071, P. R. China
  • Received:2012-03-09 Revised:2012-03-20 Published:2012-05-17
  • Contact: LIU Lian-Qing, XI Ning, ZHANG Wei-Jing E-mail:lqliu@sia.cn; xin@egr.msu.edu; zhangwj3072@163.com
  • Supported by:

    The project was supported by the National Natural Science Foundation of China (60904095, 61175103), National High Technology Research and Development Program of China (863) (2009AA03Z316), and Chinese Academy of Sciences Foreign Experts Affairs International Partnership Program for Creative Research Teams.

摘要: 原子力显微镜(AFM)的发明为研究单个活细胞的形貌结构及物理特性提供了新的技术手段. 然而, 由于缺少合适的固定方法, 利用AFM对动物悬浮活细胞的形貌进行高分辨率成像还面临着巨大的挑战. 本文提出一种基于微柱阵列和静电吸附相结合的动物悬浮细胞固定方法. 通过微柱阵列的机械钳制和多聚赖氨酸的静电吸附实现了对单个淋巴瘤B细胞的固定, 并在此基础上利用AFM动态观测了不同浓度Rituximab 刺激下淋巴瘤B细胞的表面形貌及弹性的变化. 经过0.2 mg·mL-1的Rituximab 刺激2 h 后, 细胞表面的褶皱增加, 细胞的杨氏模量从196 kPa减小到183 kPa. 经过0.5 mg·mL-1的Rituximab 刺激2 h 后, 细胞形貌发生显著变化并出现突起结构, 细胞的杨氏模量从234 kPa减小到175 kPa. 实验结果表明淋巴瘤细胞形貌和弹性变化的幅度随着Rituximab 刺激浓度的增加而增加, 加深了对Rituximab 作用效果的认识.

关键词: 原子力显微镜, 淋巴瘤, 弹性, 力曲线, 杨氏模量

Abstract: Atomic force microscopy (AFM) provides a means for characterizing the surface topography and biophysical properties of individual living cells under near-physiological conditions. However, owing to the lack of adequate cellular immobilization methods, AFM imaging of living, suspended mammalian cells is still a big challenge. In this paper, a method is presented for immobilizing individual living B lymphoma cells that combines mechanical trapping with pillar arrays and electrostatic adsorption with poly-L-lysine. In this way, the topography and elasticity changes of individual B lymphoma cells that were stimulated with different concentrations of Rituximab were observed and measured dynamically. When the cell is stimulated by 0.2 mg·mL-1 Rituximab for 2 h, the cell topography becomes more corrugated and Young's modulus decreases from 196 to 183 kPa. When the cell is stimulated by 0.5 mg·mL-1 Rituximab for 2 h, the cell topography changes more significantly and some tubercles appear, and Young's modulus decreases from 234 to 175 kPa. These results thus provide a unique insight into the effects of Rituximab on individual cells.

Key words: Atomic force microscopy, Lymphoma, Elasticity, Force curve, Young's modulus

MSC2000: 

  • O641