本文選題：表面增強拉曼光譜　切入點：細(xì)胞　出處：《福建師范大學(xué)》2011年碩士論文

【摘要】：表面增強拉曼光譜(SERS)在生物醫(yī)學(xué)領(lǐng)域具有非常廣闊的應(yīng)用前景,本論文跟蹤國際熱點研究問題,結(jié)合我國研究現(xiàn)狀和臨床需求,基于SERS技術(shù)進行鼻咽癌與胃癌檢測方法研究,主要研究成果如下： 1)分別利用細(xì)胞內(nèi)吞法、細(xì)胞自還原法以及細(xì)胞電穿孔法三種金屬納米粒子傳輸法進行細(xì)胞SERS光譜研究并對三種方法的優(yōu)缺點進行比較。重點討論利用電穿孔方法實現(xiàn)金屬納米粒子到細(xì)胞的快速輸送；首次利用電穿孔技術(shù)將銀納米粒子快速導(dǎo)入鼻咽癌C666細(xì)胞系、皮膚癌A431細(xì)胞系及白血病細(xì)胞CA46細(xì)胞系,并成功測得細(xì)胞內(nèi)SERS信號。結(jié)合電穿孔導(dǎo)入技術(shù)及全細(xì)胞SERS檢測方法,不僅可以顯著縮短細(xì)胞SERS光譜的準(zhǔn)備和測試時間,而且很大程度上提高了細(xì)胞SERS光譜的重復(fù)性和穩(wěn)定性。細(xì)胞電穿孔-SERS這一新方法將在生物醫(yī)學(xué)應(yīng)用中(如細(xì)胞內(nèi)生化物質(zhì)的實時檢測、癌細(xì)胞篩查等)展現(xiàn)出極大的應(yīng)用潛力。 2)利用金納米粒子為增強基底獲取人正常鼻咽組織與鼻咽癌組織：SERS光譜與圖像,并進行組織SERS光譜歸屬分析;探討在組織層面上觀察到的SERS光譜閃爍現(xiàn)象,研究發(fā)現(xiàn)組織SERS波動現(xiàn)象與激發(fā)光功率密度高度相關(guān)。利用SERS成像技術(shù)研究正常與癌變鼻咽組織在生化成分上的差異；在此基礎(chǔ)上進一步構(gòu)建EB病毒SERS免疫探針,證實該免疫SERS探針可實現(xiàn)高效的EB病毒免疫檢測。 3)首次提出利用血漿SERS光譜檢測實現(xiàn)非標(biāo)記鼻咽癌的無損診斷分析；利用表面增強拉曼光譜技術(shù)進行人體血漿生化分析,通過譜峰指認(rèn)進一步探討鼻咽癌患者與健康人血漿SERS光譜的差異。人體血漿SERS光譜結(jié)合主成分分析和線性判別統(tǒng)計分析可區(qū)分正常人與鼻咽癌患者血漿,診斷特異性與靈敏度分別達到100%與90.7%(針對43例鼻咽癌患者和33例正常人血漿樣品分析結(jié)果).通過對比研究發(fā)現(xiàn)鼻咽癌患者血漿與正常健康人血漿之間存在某些特定的生化成分差異。研究結(jié)果表明,血漿表面增強拉曼光譜分析很有希望發(fā)展為一種無損檢測與篩查鼻咽癌的臨床診斷工具。 4)首次利用四種不同偏振態(tài)激光激發(fā)血漿表面增強拉曼光譜并結(jié)合多變量統(tǒng)計分析方法進行胃癌的無損檢測研究。與使用非偏振激光和線偏振激光相比,利用圓偏振態(tài)激發(fā)光能夠大大提高診斷的特異性與靈敏度。利用左旋圓偏振激光進行胃癌患者血漿SERS光譜無損診斷分析,最佳診斷效果可以實現(xiàn)靈敏度為100%,特異性為97%(針對32例胃癌患者和33例正常人血漿樣品分析結(jié)果)；而利用非偏振激光激發(fā),其診斷靈敏度與特異性大概都只有72%.診斷效果的大幅度提高可能是由于左旋圓偏振激光比較適合于探測與胃癌有關(guān)的血漿手性分子。 5)首次將膜電泳提純血漿蛋白技術(shù)與表面增強拉曼光譜相結(jié)合應(yīng)用于無損癌癥檢測分析。與未經(jīng)過任何處理的血漿直接與銀膠混合后進行SERS檢測相比,這種膜電泳與SERS相結(jié)合方法可大大減少由于癌癥患者血漿中含有的外源性物質(zhì)以及除白蛋白和球蛋白之外的其它血漿成分所引起的光譜干擾變化。將血漿膜電泳蛋白SERS光譜進行主成分分析,統(tǒng)計結(jié)果顯示無論是胃癌樣本還是鼻咽癌樣本都可準(zhǔn)確地與健康樣本組區(qū)分開,診斷的靈敏度和特異性都達到100%(針對31例胃癌患者、43例鼻咽癌患者以及33例正常人血漿樣品分析結(jié)果)。我們發(fā)明的這種新方法可能會對所有類型的癌癥表現(xiàn)出敏感性,并可能成為一種非侵入性的癌癥檢測通用篩查方法。
[Abstract]:Surface - enhanced Raman spectroscopy ( SERS ) has a very wide application prospect in the field of biomedical field . This paper tracks international hotspot research , combines the current situation and clinical demand of our country , and studies the detection methods of nasopharyngeal carcinoma and gastric cancer based on SERS technique . The main research results are as follows :

1 ) The cell SERS spectra of three kinds of metal nanoparticles were studied by means of endocytosis , self - reduction and electroporation . The advantages and disadvantages of the three methods were compared .
In the first time , silver nanoparticles were rapidly introduced into NPC C666 cell line , skin cancer A431 cell line and leukemia cell CA46 cell line by electroporation technique , and SERS signal was successfully measured . Combined with electroporation and full cell SERS assay , not only the preparation and testing time of SERS spectra were significantly shortened , but also the repeatability and stability of SERS spectra were greatly improved .

2 ) using gold nanoparticles as the enhancement substrate to obtain the human normal nasopharyngeal tissue and the nasopharyngeal carcinoma tissue : SERS spectra and images , and carrying out SERS spectrum attribution analysis ; and exploring the SERS spectrum flicker phenomenon observed at the tissue level , and researching the difference of the tissue SERS fluctuation phenomenon and the excitation light power density .
On the basis of this , the EB virus SERS immunoprobe was further constructed , and it was confirmed that the immune SERS probe could realize high - efficiency EB virus immunodetection .

3 ) A nondestructive diagnostic analysis of non - labeled nasopharyngeal carcinoma ( NPC ) using plasma SERS ( SERS ) was proposed for the first time .
The plasma SERS spectra of patients with nasopharyngeal carcinoma ( NPC ) and normal healthy human plasma were analyzed by means of surface enhanced Raman spectroscopy ( SERS ) . The results showed that the plasma surface enhanced Raman spectrum analysis was promising to develop a clinical diagnostic tool for non - destructive testing and screening of nasopharyngeal carcinoma .

4 ) Using four different polarization states laser to excite plasma surface enhanced Raman spectrum and combined with multivariate statistical analysis method to study the non - destructive examination of gastric cancer . Compared with non - polarized laser and linear polarized laser , using circular polarized laser to excite light can greatly improve the specificity and sensitivity of diagnosis .
With non - polarized laser excitation , the diagnostic sensitivity and specificity can be only 72 % . The significant increase in diagnostic effectiveness may be due to the comparison of the left - handed circular polarized laser to the detection of the plasma chiral molecules associated with gastric cancer .

The plasma membrane electrophoretic protein SERS ( SERS ) spectra were analyzed by SERS spectra . The results showed that the sensitivity and specificity of plasma membrane electrophoresis were 100 % ( for 31 patients with gastric cancer , 43 patients with nasopharyngeal carcinoma and 33 normal human plasma samples ) . This new method of our invention may show sensitivity to all types of cancer and may be a non - invasive cancer detection universal screening method .

【學(xué)位授予單位】：福建師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R739.63;R735.2;O657.37

【引證文獻】

相關(guān)碩士學(xué)位論文 前1條

1 陳永堅;基于拉曼光譜的茶氨酸研究及茶葉種類判別分析[D];福建師范大學(xué);2012年

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本文編號：1678256