Traditional technique such nanoindenter（NI） can＇t measure the local elastic modulus at nano-scale（lateral）. Atomic force acoustic microscopy （AFAM） is a dynamic method, which can quantitatively determine indent...Traditional technique such nanoindenter（NI） can＇t measure the local elastic modulus at nano-scale（lateral）. Atomic force acoustic microscopy （AFAM） is a dynamic method, which can quantitatively determine indentation modulus by measuring the contact resonance spectra for high order modes of the cantilever. But there are few reports on the effect of experimental factors, such length of cantilever, contact stiffness on measured value. For three different samples, including copper（Cu） film with 110 nm thickness, zinc（Zn） film of 90 nm thickness and glass slides, are prepared and tested, using referencing approach in which measurements are performed on the test and reference samples （it＇s elastic modulus is known）, and their contact resonance spectra are measured used the AFAM system experimentally. According to the vibration theory, from the lowest two contact resonance frequencies, the tip-sample contact stiffness is calculated, and then the values for the elastic properties of test sample, such as the indentation modulus, are determined. Using AFAM system, the measured indentation modulus of copper nano-film, zinc nano-film and glass slides are 113.53 GPa, 87.92 GPa and 57.04 GPa, which are agreement with literature values Mcu--105-130 GPa, Mzn = 88.44 GPa and Molass = 50-90 GPa. Furthermore, the sensitivity of contact resonance frequency to contact stiffness is analyzed theoretically. The results show that for the cantilevers with the length 160 pm, 225 μm and 520 μm respectively, when contact stiffness increases from 400 N/m to 600 N/m, the increments of first contact resonance frequency are 126 kHz, 93 kHz and 0.6 kHz, which show that the sensitivity of the contact resonance frequency to the contact stiffness reduces with the length of cantilever increasing. The novel method presented can characterize elastic modulus of near surface for nano-film and bulk material, and local elasticity of near surface can be evaluated by optimizing the experimental parameters using the AFAM system.展开更多
目的:采用原子力声显微镜及透射电镜观察低频低能量超声联合微泡对前列腺癌细胞DU145及正常前列腺上皮细胞RWPE-1的作用。方法:两种细胞均分为对照组、单纯超声组、超声联合微泡组。对照组加入一定比例的生理盐水,不进行超声辐照;单纯...目的:采用原子力声显微镜及透射电镜观察低频低能量超声联合微泡对前列腺癌细胞DU145及正常前列腺上皮细胞RWPE-1的作用。方法:两种细胞均分为对照组、单纯超声组、超声联合微泡组。对照组加入一定比例的生理盐水,不进行超声辐照;单纯超声组加入相同比例的生理盐水,用发射频率为21 k Hz的低频超声辐照,辐照2 min,占空比30%;超声联合微泡组加入同前相同比例的微泡造影剂悬浊液,用与单纯超声组相同的超声辐照。处理过的细胞立即用原子力声显微镜观察其形貌并计算其杨氏模量。同时,对同一处理方式的对照组及超声联合微泡组细胞继续培养24 h后,用透射电镜观察细胞。结果:单纯超声组的DU145细胞及RWPE-1细胞的细胞形态与对照组无明显变化;超声联合微泡组的DU145细胞及RWPE-1细胞形态呈类圆形,细胞表面可见放射状显微丝状结构,细胞膜表面可见多个大孔状结构;超声联合微泡组的DU145细胞的弹性模量较RWPE-1细胞大,且与对照组相比,两种细胞的弹性模量均变大,DU145细胞尤甚。透射电镜观察结果示对照组的两种细胞未见细胞自噬,而超声联合微泡组两种细胞都出现自噬现象,其中DU145细胞中可见凋亡现象。结论:低频低能量超声联合微泡可引起前列腺癌DU145细胞及正常前列腺上皮RWPE-1细胞的细胞膜出现孔状结构,并诱导其自噬,且对DU145细胞的损伤大于RWPE-1细胞。展开更多
基金supported by National Natural Science Foundation of China(Grant No. 50775005)General Program of Science and Technology Development Project of Beijing Municipal Education Commission(Grant No. KM201110015009)
文摘Traditional technique such nanoindenter（NI） can＇t measure the local elastic modulus at nano-scale（lateral）. Atomic force acoustic microscopy （AFAM） is a dynamic method, which can quantitatively determine indentation modulus by measuring the contact resonance spectra for high order modes of the cantilever. But there are few reports on the effect of experimental factors, such length of cantilever, contact stiffness on measured value. For three different samples, including copper（Cu） film with 110 nm thickness, zinc（Zn） film of 90 nm thickness and glass slides, are prepared and tested, using referencing approach in which measurements are performed on the test and reference samples （it＇s elastic modulus is known）, and their contact resonance spectra are measured used the AFAM system experimentally. According to the vibration theory, from the lowest two contact resonance frequencies, the tip-sample contact stiffness is calculated, and then the values for the elastic properties of test sample, such as the indentation modulus, are determined. Using AFAM system, the measured indentation modulus of copper nano-film, zinc nano-film and glass slides are 113.53 GPa, 87.92 GPa and 57.04 GPa, which are agreement with literature values Mcu--105-130 GPa, Mzn = 88.44 GPa and Molass = 50-90 GPa. Furthermore, the sensitivity of contact resonance frequency to contact stiffness is analyzed theoretically. The results show that for the cantilevers with the length 160 pm, 225 μm and 520 μm respectively, when contact stiffness increases from 400 N/m to 600 N/m, the increments of first contact resonance frequency are 126 kHz, 93 kHz and 0.6 kHz, which show that the sensitivity of the contact resonance frequency to the contact stiffness reduces with the length of cantilever increasing. The novel method presented can characterize elastic modulus of near surface for nano-film and bulk material, and local elasticity of near surface can be evaluated by optimizing the experimental parameters using the AFAM system.
文摘目的:采用原子力声显微镜及透射电镜观察低频低能量超声联合微泡对前列腺癌细胞DU145及正常前列腺上皮细胞RWPE-1的作用。方法:两种细胞均分为对照组、单纯超声组、超声联合微泡组。对照组加入一定比例的生理盐水,不进行超声辐照;单纯超声组加入相同比例的生理盐水,用发射频率为21 k Hz的低频超声辐照,辐照2 min,占空比30%;超声联合微泡组加入同前相同比例的微泡造影剂悬浊液,用与单纯超声组相同的超声辐照。处理过的细胞立即用原子力声显微镜观察其形貌并计算其杨氏模量。同时,对同一处理方式的对照组及超声联合微泡组细胞继续培养24 h后,用透射电镜观察细胞。结果:单纯超声组的DU145细胞及RWPE-1细胞的细胞形态与对照组无明显变化;超声联合微泡组的DU145细胞及RWPE-1细胞形态呈类圆形,细胞表面可见放射状显微丝状结构,细胞膜表面可见多个大孔状结构;超声联合微泡组的DU145细胞的弹性模量较RWPE-1细胞大,且与对照组相比,两种细胞的弹性模量均变大,DU145细胞尤甚。透射电镜观察结果示对照组的两种细胞未见细胞自噬,而超声联合微泡组两种细胞都出现自噬现象,其中DU145细胞中可见凋亡现象。结论:低频低能量超声联合微泡可引起前列腺癌DU145细胞及正常前列腺上皮RWPE-1细胞的细胞膜出现孔状结构,并诱导其自噬,且对DU145细胞的损伤大于RWPE-1细胞。
