【作者】 吴志超；

【导师】 王景贺；

【作者基本信息】 哈尔滨工业大学 ， 机械工程， 2011， 硕士

【摘要】 癌症,近几十年来已对人类健康构成强大威胁,虽然对于肿瘤的组织病理学诊断和放化疗技术日趋成熟,但癌症的发病机理和转移机制等根本性问题却尚未明晰。利用AFM在单细胞水平对癌细胞进行相关分析已成为当今癌症研究的最新热点,具有其它技术无法比拟的优势,但大多集中于AFM形貌成像等基础研究,从机械角度出发对于癌细胞的力学特性进行分析限于研究手段等原因却受到较少关注,而力学特性与癌细胞转移过程息息相关。本课题正是利用自行制作的小球探针,基于AFM的Nanoindentation功能在液态环境下对癌细胞的力学特性进行测定,分析其对转移的影响。首先,对现有的应用于固体纳米压的痕三角形悬臂探针针尖进行功能化改造,扩展了AFM在生物领域的应用。利用实验室现有设备,自行制作出满足液态下对活细胞进行纳米压痕的小球探针,并利用Cleveland方法对其弹性常数进行了校准。该制作过程同样适用于其它微米级球状物向微米级尖状物的粘附。然后,分析了液态环境下实施活细胞纳米压痕实验所涉及的各方面条件,包括活细胞的选择、培养、转运,液态实验环境建立,仪器的改装,实验的改进,操作的优化,以及具体实验前仪器的标定等等,每一项都关乎实验能否顺利进行。其中自行总结的液态下压痕目标细胞快速定位方法具有很好的实用性和通用性。最后,在保持细胞最佳生理活性的液态环境下,对转移性不同的Anip-973和AGZY-83a两种肺癌细胞进行AFM纳米压痕实验,并利用Hertz接触方程及应力松弛模型对实验结果进行非线性拟合,再通过方差分析比对力学特性参数结果,以研究其对癌变转移的影响。更多还原

【Abstract】 Cancer has posed a powerful threat to human health in recent decades. The histopathological diagnosis and chemotherapy technology of the tumor is maturing; however, fundamental issues as the pathogenesis of cancer and metastatic mechanism stay vague. As the latest hot spot researching cancer cells in the single-cell level based AFM, the technology can not match advantage, but most focused on basic research, e.g. the AFM topography imaging. The method analyzing mechanical properties of cancer cells from a mechanical view has been limited by investigating way, which has been less reason for concern, are closely related with cancer cells metastasis. The paper aimed to research their mechanical properties and the impact of metastasis in the use of self-made spherical ball tip, based on the AFM nanoindentation features in the liquid environment.First, functional transformation of existing solid triangle cantilever probe tip used in solid namoindentation, extended the application of AFM in the biological field. By laboratory equipment existing, fabricated spherical tip fitting for nanoindentation of living cells in liquid by self, and calibrated its elastic constants by Cleveland method. The production process also applies to stick other micron ball to the micron spikes.Then, analyzed of all aspects of the conditions involved in the nanoindentation of living cells in the liquid environment, including the choice of living cells, culture, transport, liquid experiment environment to establish, what’s more, the modification of the equipment, test improvement, operation optimization, and the specific calibration of the instrument before the experiment, etc., each of which relates to experiment can proceed smoothly. The summary of quick cell targeting in liquid indentation has good practicality and versatility.Finally, in the PBS liquid environment to maintain their best physical activity, experiments were performed between Anip-973 and AGZY-83a lung cancer cells with different metastatic ability. The results were implemented nonlinear fitting upon the stress-relaxation model derived from Hertz contact equation. ANOVA was conducted to determine the differences between each of the mechanical parameters, in order to examine its impact on cancer metastasis.更多还原