人肺鳞癌细胞SK-MES-1介电频谱特性研究

发布时间：2018-03-28 17:16

本文选题：介电谱　切入点：肺鳞癌细胞　出处：《宁波大学》2015年硕士论文

【摘要】：目的:作为一项成熟的,具备完善测量手段和分析方法的频谱技术,介电谱的应用范围已经从最初的物理学领域扩展到生物学和医学等学科。本论文以细胞悬浮液中人肺癌细胞的生物学变化对应的介电响应为主要研究内容,探讨将生物介电谱测量和分析技术应用到人肺癌症筛查,诊断,以及癌细胞分型等临床需求中的可行性。方法:本论文首先在100 Hz~100 MHz交变电场频率范围内,使用安捷伦4294A阻抗分析仪和两电极的测量池,对人正常肺上皮细胞BEAS-2B和人肺鳞癌细胞SK-MES-1两种细胞不同细胞体积分数的悬浮液进行介电测量。然后结合三项式Cole-Cole方程和单壳物理模型对测量得到的介电谱进行分析,应用非线性最小二乘曲线拟合技术提取ColeCole模型参数和细胞形态学参数。最后通过分析BEAS-2B和SK-MES-1的介电谱特征参数和解析模型参数差异,探讨在实验条件下实现肺癌细胞与正常细胞分型的可行性和具体实施细节。本论文还尝试从生物物理学角度为人肺鳞癌细胞的病理学研究提供辅助信息,并结合细胞电镜和光镜照片讨论介电谱差异与细胞形态学变化之间的关系。结果:1肺鳞癌细胞的介电常数较正常细胞的介电常数低(5.81%),但肺鳞癌细胞的电导率较正常细胞的电导率高(12.25%)。2 BEAS-2B和SK-MES-1细胞悬浮液的介电谱、电导率谱、复平面图和介电损耗谱均表现有细胞悬浮液体积分数依存性。3 BEAS-2B和SK-MES-1细胞悬浮液在100 Hz~100 MHz频率范围均包含α和β两个介电弛豫。4对BEAS-2B和SK-MES-1细胞悬浮液的介电谱进行Cole-Cole模型分析,需要使用三项式Cole-Cole方程以获得满足精度要求的拟合结果。这表明两类细胞悬浮液的介电谱均对应三个特征频率,分别表征了不同频率段的外加电场激发产生的双电层中对离子极化,以及细胞膜和核膜处的界面极化。5应用Cole-Cole模型分析细胞悬浮液的介电谱得到的三个特征频率(fc1,fc2,fc3)不受细胞体积分数的影响。统计学分析结果显示特征频率可以作为区分BEAS-2B和SKMES-1的依据。6应用单壳球形细胞物理模型对两类细胞的介电谱进行解析,得到的物理模型参数差异不仅可以体现两细胞的介电谱差异,而且结合两细胞的电镜和光镜照片结果,可以解释细胞癌变后形态学变化与介电差异的关系。结论:本论文的结果表明人正常肺上皮细胞BEAS-2B和人肺鳞癌细胞SK-MES-1的介电谱具有明显差异。基于Cole-Cole模型和物理模型,结合曲线拟合技术可以提取特征频率等模型参数,特征化了两类细胞的介电谱差异,可以用做细胞分型的重要依据,为基于生物介电谱技术的人肺癌细胞分型等应用提供了有力的证明。
[Abstract]:Objective: as a mature spectrum technology with perfect means of measurement and analysis, The application of dielectric spectroscopy has been extended from physics to biology and medicine. In this thesis, the dielectric response of human lung cancer cells in cell suspensions is the main research content. To explore the feasibility of applying biological dielectric spectrum measurement and analysis technology to the clinical requirements of human lung cancer screening, diagnosis, and cancer cell typing. Methods: in this paper, the frequency of 100 Hz~100 MHz AC electric field was first measured. Using the Agilent 4294A impedance analyzer and a two-electrode measuring cell, The dielectric properties of human normal lung epithelial cells (BEAS-2B) and human lung squamous cell carcinoma (SK-MES-1) cells with different cell volume fraction were measured. The dielectric spectra were analyzed by using trinomial Cole-Cole equation and single-shell physical model. The nonlinear least square curve fitting technique is used to extract the parameters of ColeCole model and cell morphology. Finally, the difference of dielectric spectrum characteristic parameters and analytical model parameters between BEAS-2B and SK-MES-1 is analyzed. To explore the feasibility and practical details of the classification of lung cancer cells from normal cells under experimental conditions. This paper also attempts to provide auxiliary information for the pathological study of human lung squamous cell carcinoma cells from the perspective of biophysics. The relationship between the difference of dielectric spectrum and the morphological changes of lung squamous cell carcinoma cells was discussed by means of electron microscope and light microscope. Results the dielectric constant of lung squamous cell carcinoma cells was lower than that of normal cells by 5.81%, but the electrical conductivity of lung squamous cell carcinoma cells was more positive. The electrical conductivity of normal cells is high 12.25%. The dielectric spectra of BEAS-2B and SK-MES-1 cell suspensions, Conductivity spectrum, The complex plane and dielectric loss spectra showed that the cell suspension volume fraction dependence was 3. 3 BEAS-2B and the SK-MES-1 cell suspension included 伪 and 尾 in the frequency range of 100 Hz~100 MHz. The dielectric spectra of BEAS-2B and SK-MES-1 cell suspensions were analyzed by Cole-Cole model, and the dielectric spectra of BEAS-2B and SK-MES-1 cell suspensions were analyzed by Cole-Cole model. It is necessary to use the trinomial Cole-Cole equation to obtain the fitting results which meet the precision requirements. This indicates that the dielectric spectra of the two cell suspensions correspond to three characteristic frequencies. The polarization of the ions in the two-layer induced by the external electric field at different frequencies was characterized, respectively. And the interfacial polarization of cell membrane and nuclear membrane. 5. The three characteristic frequencies obtained by using Cole-Cole model to analyze the dielectric spectrum of cell suspensions were not affected by the cell volume fraction. The results of statistical analysis showed that the characteristic frequency could be used as a kind of characteristic frequency. The dielectric spectra of BEAS-2B and SKMES-1 were analyzed by using the physical model of single-shell spherical cells. The difference of physical model parameters can not only reflect the difference of dielectric spectrum of two cells, but also combine with the results of electron microscope and light microscope photos of the two cells. Conclusion: the results of this paper indicate that there are significant differences in dielectric spectra between human normal lung epithelial cells (BEAS-2B) and human lung squamous cell carcinoma cells (SK-MES-1) based on Cole-Cole model and physical model. The characteristic frequency and other model parameters can be extracted by the curve fitting technique, and the dielectric spectrum difference between the two kinds of cells can be characterized, which can be used as an important basis for cell typing. It provides a powerful proof for the application of human lung cancer cell typing based on biological dielectric spectroscopy.
【学位授予单位】：宁波大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R734.2

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