拉曼光谱技术在肿瘤鉴别与异物定性中的应用研究

发布时间：2018-06-21 03:08

本文选题：拉曼光谱 + CTC　；参考：《济南大学》2016年硕士论文

【摘要】：目的结合拉曼光谱技术在生物医学中的应用情况,本实验拟应用拉曼光谱技术鉴别课题组迪夫染色的循环肿瘤细胞(Circulating Tumor Cells,CTCs)与外周血淋巴细胞;鉴别瑞氏-姬姆萨染色的肿瘤细胞与正常细胞;鉴别肝癌细胞系与外周血淋巴细胞;鉴别肝癌癌组织与合并肝硬化肝组织以及鉴定膈肌异物。方法本文运用拉曼光谱技术在细胞层面分析了迪夫染液样本中CTCs、CTCs旁淋巴细胞及样本中滤膜、染料、玻片等的拉曼光谱;瑞氏-姬姆萨染色样本中的肿瘤细胞与正常细胞的拉曼光谱;肝癌细胞系与正常人外周血白细胞的拉曼光谱。在组织层面分析了30例肝癌手术患者的肝癌癌组织与合并肝硬化肝组织的拉曼光谱。在临床案例中分析了肿物中异物的拉曼光谱。结果通过分析各样本拉曼光谱,结果显示:1)在细胞层面,532 nm波长激光下经迪夫染色的CTCs、淋巴细胞及染料的拉曼光谱在209~213 cm-1、273~278 cm-1、336~338 cm-1等峰位处都有类似特征峰;785 nm波长激光下经迪夫染色的肿瘤细胞、滤膜与玻片的拉曼光谱在1200~2000 cm-1区域都存在高耸的荧光峰,在1200 cm-1以前区域,631~632cm-1、701~702 cm-1、883~887 cm-1、1106~1107 cm-1处鼻咽癌细胞与滤膜有拉曼特征峰,而空白玻片呈光滑曲线。在532nm波长激光下经瑞氏-姬姆萨染色的肿瘤细胞与淋巴细胞在208~215 cm-1、278~285 cm-1、336~341 cm-1等峰位附近都有类似特征峰而染料由于染色较浅除在峰位640~643 cm-1、708~713 cm-1、1500~1505 cm-1、1620~1624cm-1处拉曼特征峰峰强较高外,其余峰强均较低。白细胞与细胞周围基底的拉曼光谱曲线平缓,没有显著的特征峰;而肝癌细胞在峰位1003 cm-1、1449 cm-1、1666 cm-1附近可见显著特征峰。2)组织层面,从肝癌癌组织与合并肝硬化肝组织的拉曼光谱对比图可以看出:在峰位1004 cm-1、1155 cm-1、1510 cm-1附近合并肝硬化肝组织的峰强高于肝癌癌组织;在峰位1642 cm-1附近合并肝硬化肝组织的峰强低于肝癌癌组织;在1377 cm-1和1586 cm-1附近肝癌癌组织有显著特征峰而合并肝硬化的肝组织没有。3)在临床案例中,从膈肌异物、中人氟安及氟尿嘧啶注射液的拉曼光谱可以看出在412cm-1、687 cm-1、1344 cm-1、1538 cm-1、2876 cm-1等处都有明显特征峰且高度大致相同。结论通过拉曼光谱技术分析迪夫染色样本与瑞氏-姬姆萨染色样本的拉曼光谱可见通过染色后的样本由于染料等因素的干扰用拉曼光谱技术是无法鉴别的,通过对未染色样本肝癌细胞系与人外周血淋巴细胞拉曼光谱的分析可见运用拉曼光谱技术鉴别肿瘤细胞与淋巴细胞是可行的。通过对30例肝癌癌组织与合并肝硬化肝组织拉曼光谱的对比可以看出,运用拉曼光谱技术鉴别肝癌癌组织与合并肝硬化的肝组织是可行的。通过拉曼光谱对临床工作中异物性质的确定,为我们临床工作对异物的鉴定提供了一种新手段。
[Abstract]:Objective to identify circulating tumor cells (CTCss) and peripheral blood lymphocytes (PBL) from circulating tumor cells (CTCss) by using Raman spectroscopy in biomedicine. The tumor cell lines and peripheral blood lymphocytes were differentiated between tumor cells and normal cells stained by Ricker-Giemsa staining, liver tissues and liver tissues with liver cirrhosis and diaphragmatic foreign bodies were identified by distinguishing liver cancer cell lines from peripheral blood lymphocytes. Methods Raman spectroscopy was used to analyze the Raman spectra of the lymphocytes in CTCs and the filter membrane, dyes and glass slides in the samples of Duff dye solution at the cell level. The Raman spectra of tumor cells and normal cells in the samples stained by Rish-Jimsa, and the Raman spectra of leukocytes in peripheral blood of hepatoma cells and normal people. The Raman spectra of liver cancer tissues and liver tissues associated with liver cirrhosis in 30 patients undergoing liver cancer surgery were analyzed at the tissue level. Raman spectra of foreign bodies in tumor were analyzed in clinical cases. Results by analyzing the Raman spectra of each sample, The results showed that the Raman spectra of lymphocytes and dyes at the wavelength of 532nm at cell level had a similar characteristic peak at the peak position of 209 ~ 213 cm-1273~278 cm-1336~338 cm-1. The Raman spectra of the filter membrane and the glass plate have high fluorescence peaks in the region of 1 200 ~ 2 000 cm-1. In the region before 1200 cm-1, there is a Raman characteristic peak between nasopharyngeal carcinoma cells and the filter membrane in the region of 631C ~ 632 cm ~ (-1) ~ (-1) ~ 702 cm-1883~887 ~ (-1) C ~ (-1) ~ (-1) ~ 1106C ~ (-1) cm-1, while the blank glass sheet shows a smooth curve. Under 532nm wavelength laser, the tumor cells and lymphocytes stained by Rayleigh Giemsa have similar characteristic peaks near the isotherm position of 208 ~ 215 cm-1278~285 cm-1336~341 cm-1. The intensity of Raman characteristic peaks is higher than that at 640643 cm-1708~713 cm ~ (-1) 1505 cm ~ (-1) and 1620 ~ (-1) cm ~ (-1) due to the dye staining, but the intensity of Raman peaks at 640643 cm-1708~713 / cm ~ (-1) and 1620 ~ (-1) cm ~ (-1) is higher than that at 640 ~ 643 cm-1708~713 / cm ~ (-1). The other peaks were lower. The Raman spectra of white blood cells and their surrounding substrates were smooth, and there was no significant characteristic peak, while that of hepatoma cells was 1003 cm ~ (-1) ~ 1449 cm ~ (-1) ~ 1666 cm-1. From the Raman spectra of liver cancer tissue and liver tissue with liver cirrhosis, it can be seen that the peak intensity of liver tissue with liver cirrhosis is higher than that of liver cancer tissue at the peak of 1004 cm ~ (-1) ~ 1155 cm ~ (-1) ~ 1510 cm-1. The peak intensity of liver tissue associated with liver cirrhosis was lower than that of liver cancer tissue in the vicinity of peak position 1642 cm-1, and that of liver cancer tissue with liver cirrhosis in the vicinity of 1377 cm-1 and 1586 cm-1 was significantly higher than that of liver tissue with liver cirrhosis. The Raman spectra of human fluorine and fluorouracil injection show that there are obvious characteristic peaks at 412cm-1687 cm-1 + 1344cm-1n 1538cm-1n 2876 cm-1 and the height is about the same. Conclusion the Raman spectra of the dyed samples and the samples stained by Reich Giemsa can not be distinguished by Raman spectroscopy because of the interference of dye and other factors. The Raman spectra of unstained hepatoma cell lines and human peripheral blood lymphocytes showed that it was feasible to identify tumor cells and lymphocytes by Raman spectroscopy. By comparing the Raman spectra of 30 cases of liver cancer with liver cirrhosis, we can see that it is feasible to use Raman spectroscopy to distinguish liver tissues from liver cancer and liver cirrhosis. The determination of the properties of foreign bodies in clinical work by Raman spectroscopy provides a new method for the identification of foreign bodies in clinical work.
【学位授予单位】：济南大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.7

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