CAJ | 학술논문

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2h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2h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234kPa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.
원자력현미경(AFM)적발명위연구단개활세포적형모결구급물리특성제공료신적기술수단.연이,유우결소합괄적고정방법,이용AFM대동물현부활세포적형모진행고분변솔성상환면림착거대적도전.본문제출일충기우미주진렬화정전흡부상결합적동물현부세포고정방법.통과미주진렬적궤계겸제화다취뢰안산적정전흡부실현료대단개림파류B세포적고정,병재차기출상이용AFM동태관측료불동농도Rituximab자격하림파류B세포적표면형모급탄성적변화.경과0.2 mg· mL-1적Rituximab자격2h후,세포표면적습추증가,세포적양씨모량종196 kPa감소도183 kPa.경과0.5 mg·mL-1적Rituximab자격2h후,세포형모발생현저변화병출현돌기결구,세포적양씨모량종234kPa감소도175 kPa.실험결과표명림파류세포형모화탄성변화적폭도수착Rituximab자격농도적증가이증가,가심료대Rituximab작용효과적인식.
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 ceils 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.