本文選題：生物傳感器　切入點(diǎn)：場(chǎng)效應(yīng)晶體管　出處：《湖北中醫(yī)藥大學(xué)》2017年碩士論文

【摘要】：一氧化氮既是信使分子又是神經(jīng)遞質(zhì),可以接收和傳達(dá)信息以調(diào)節(jié)細(xì)胞活性,并且指示機(jī)體執(zhí)行某些重要功能,例如神經(jīng)傳遞、血管舒張、免疫應(yīng)答和血管生成等。研究發(fā)現(xiàn)一氧化氮不但在動(dòng)物的免疫調(diào)節(jié)、神經(jīng)元通訊、平滑肌松弛等活動(dòng)中充當(dāng)信號(hào)分子,而且參與植物的生長(zhǎng)發(fā)育、脅迫反應(yīng)、呼吸代謝以及衰老成熟等生理過(guò)程。然而,一氧化氮作為活性氣體在體內(nèi)以納摩爾濃度存在,具有相對(duì)短的半衰期(10s),且極易被氧化物或超氧化物離子氧化,使得難以精確地量化生物系統(tǒng)中的一氧化氮水平,因此,高效、準(zhǔn)確、定量獲取細(xì)胞及體內(nèi)一氧化氮信號(hào)分子信息則是深入探索其作用機(jī)制的基礎(chǔ)。到目前為止,許多用于一氧化氮檢測(cè)的技術(shù)被應(yīng)用,例如生物測(cè)定,電子磁共振光譜,魯米諾/過(guò)氧化氫化學(xué)發(fā)光,激光誘導(dǎo)熒光,分光光度測(cè)定等。然而,這些方法還存在成本高、預(yù)處理復(fù)雜和靈敏度低等缺點(diǎn)。電化學(xué)生物傳感器作為一種有效的檢測(cè)工具,可以高靈敏度、實(shí)時(shí)監(jiān)測(cè)一氧化氮的釋放,已被廣泛用于生物系統(tǒng)中一氧化氮的檢測(cè)。然而,為實(shí)現(xiàn)一氧化氮檢測(cè)的目的,電化學(xué)一氧化氮傳感器通常需要在相對(duì)高的氧化電位(0.6V)下操作,這會(huì)干擾固有的細(xì)胞反應(yīng)或誘導(dǎo)來(lái)自其它電化學(xué)活性分子顯著的信號(hào)干擾。與安培計(jì)電極相比,場(chǎng)效應(yīng)晶體管納米生物傳感器由于其高信噪比可實(shí)施高靈敏、實(shí)時(shí)的監(jiān)測(cè),同時(shí),它在相對(duì)較低的檢測(cè)電位(Vds=0.1V)條件下工作,這對(duì)細(xì)胞幾乎沒(méi)有影響。因此,使用場(chǎng)效應(yīng)晶體管生物傳感器實(shí)時(shí)檢測(cè)一氧化氮具有非常重要的意義。近年來(lái),基于場(chǎng)效應(yīng)晶體管生物傳感器用于實(shí)時(shí)監(jiān)測(cè)一氧化氮的工作已經(jīng)被報(bào)道。中國(guó)科學(xué)院蘇州納米所劉立偉課題組報(bào)道了基于石墨烯的場(chǎng)效應(yīng)晶體管生物傳感器檢測(cè)一氧化氮?dú)怏w;以色列魏茨曼科學(xué)研究院Shvarts課題組報(bào)道了一種砷化鎵場(chǎng)效應(yīng)晶體管生物傳感器用于檢測(cè)生理溶液中一氧化氮。然而,這些傳感器的靈敏度仍然局限于微摩爾或亞微摩爾水平,且它們均未用于細(xì)胞水平的檢測(cè)。最近,美國(guó)南加州大學(xué)段鑲鋒課題組報(bào)道了基于血紅素功能化石墨烯場(chǎng)效應(yīng)晶體管的一氧化氮傳感器直接檢測(cè)活細(xì)胞中釋放的一氧化氮。然而,該方法未能實(shí)現(xiàn)單細(xì)胞水平一氧化氮的釋放。細(xì)胞是生物體的基本單位,在單細(xì)胞水平的檢測(cè)將增強(qiáng)對(duì)多種細(xì)胞過(guò)程如細(xì)胞內(nèi)和細(xì)胞間通訊,細(xì)胞分化,外部刺激的生理效應(yīng)和疾病狀態(tài)等的認(rèn)識(shí)。此外,對(duì)于診斷基于細(xì)胞的疾病,也需要開(kāi)發(fā)高靈敏度的方法用于單細(xì)胞水平的檢測(cè),以獲得最好的診斷能力。因此,為了實(shí)現(xiàn)單細(xì)胞水平一氧化氮釋放的實(shí)時(shí)監(jiān)測(cè),研究高靈敏度的新型場(chǎng)效應(yīng)晶體管生物傳感器非常必要。針對(duì)以上現(xiàn)狀和挑戰(zhàn),本論文立足于發(fā)展高性能場(chǎng)效應(yīng)晶體管生物傳感器以解決在單細(xì)胞水平實(shí)時(shí)監(jiān)測(cè)細(xì)胞釋放的一氧化氮。在本工作中,我們發(fā)展了一種基于還原氧化石墨烯和鐵卟啉功能化石墨烯的場(chǎng)效應(yīng)晶體管生物傳感器,用于在單細(xì)胞水平超靈敏和高效率實(shí)時(shí)監(jiān)測(cè)一氧化氮。將還原氧化石墨烯和金屬鐵卟啉通過(guò)π-π堆疊作用層層組裝在事先制備好的石墨烯場(chǎng)效應(yīng)晶體管生物傳感器溝道表面構(gòu)建卟啉功能化的石墨烯場(chǎng)效應(yīng)晶體管生物傳感器,由于石墨烯良好的導(dǎo)電性和卟啉分子對(duì)一氧化氮特異的催化性能,該傳感器能實(shí)現(xiàn)對(duì)一氧化氮超靈敏和高特異性的檢測(cè)。實(shí)驗(yàn)證明,該生物傳感器展現(xiàn)出了優(yōu)良的導(dǎo)電性和高催化性能,能夠?qū)崟r(shí)監(jiān)測(cè)1pM至100nM范圍內(nèi)的一氧化氮,檢測(cè)限可低至1pM,同時(shí)還實(shí)現(xiàn)了在單細(xì)胞水平超靈敏和高特異性實(shí)時(shí)監(jiān)測(cè)人臍靜脈內(nèi)皮細(xì)胞釋放的一氧化氮。因其具有微型化、超靈敏和快速響應(yīng)特性,該生物傳感器在生物應(yīng)用和臨床診斷方面有很大潛力。
[Abstract]:Nitric oxide is messenger molecule and neurotransmitter, can receive and transmit information to regulate cell activity, and indicates the body to perform some important functions, such as vasodilation, neurotransmission, immune response and angiogenesis. It is found that nitric oxide not only in animal immune, neuronal communication, act as signaling molecules of smooth muscle relaxation and other activities, and involved in plant growth and development, stress response, respiratory metabolism and senescence and other physiological processes. However, the activity of nitric oxide as a gas in the body at nanomolar concentrations, has a relatively short half-life (10s), and can easily be oxide or superoxide ion oxidation, making it difficult to accurately quantify the level of nitric oxide in biological systems, therefore efficient, accurate, quantitative, obtaining cells and in vivo nitric oxide signaling molecular information is to further explore the mechanism of the base The foundation. So far, many for nitric oxide detection technologies are applied, such as bioassay, electron paramagnetic resonance spectroscopy, hydrogen peroxide / luminol chemiluminescence, laser induced fluorescence, spectrophotometry and so on. However, these methods still have high cost, complicated pretreatment and low sensitivity. As an effective electrochemical biosensor the detection tool, can high sensitivity, real-time monitoring of the release of nitric oxide, nitric oxide detection has been widely used in biological systems. However, in order to achieve the purpose of electrochemical detection of nitric oxide, nitric oxide sensor usually in a relatively high oxidation potential (0.6V) under operation, it will interfere with the inherent cellular responses or induced from a signal the electrochemical activity of molecules significantly. Compared with the ammeter electrode, field effect transistor biosensors because of its high signal-to-noise ratio. Application of high sensitivity, real-time monitoring, at the same time, it is in a relatively low detection potential (Vds=0.1V) under the working conditions, almost no effect on the cells. Therefore, has very important significance to use field effect transistor biosensor for real-time detection of nitric oxide. In recent years, based on field effect transistor biosensor for real-time monitoring of nitric oxide the work has been reported. China Academy of Sciences Suzhou Institute for nanotechnology research group reported the Liu Liwei gas field effect transistor biosensor based on graphene oxide; Israel Weizmann Scientific Research Institute of the Shvarts research group of a GaAs FET biosensor for detection of nitric oxide in physiological solution has been reported. However, the sensitivity of these sensors is limited to micromolar or submicromolar levels, and they were not used to detect cell level. Recently, the United States and South Section Xiangfeng State University research group reported a direct detection of nitric oxide release in living cells heme functionalized graphene field effect transistor nitric oxide sensor based on single cell level. However, the release of nitric oxide. The method failed to cell is the basic unit of organism, detection at the single cell level will be enhanced to a variety of cellular processes such as cell intra - and intercellular communication, cell differentiation, understanding the physiological effects of external stimuli and disease status. In addition, the cells in the diagnosis of disease based on the method, but also need to develop a high sensitivity for the detection of single cell level, in order to obtain the best diagnostic ability. Therefore, in order to realize the real-time monitoring of single cell level of nitric oxide release. Study on the high sensitivity of the new field effect transistor biosensor is very necessary. In view of the above situation and challenges, this paper is based on the development of high The performance of field effect transistor biosensor in order to solve the problems in real-time monitoring of nitric oxide release in single cell level. In this work, we develop a reduction of graphene oxide and iron porphyrin functionalized graphene field effect transistor biosensor based on single cell level for ultra sensitive and efficient real-time monitoring of graphene oxide nitric oxide. Graphene and metal iron porphyrin reduction by tt-tt stacking layers assembled in the prepared graphene field effect transistor biosensor channel surface construction of porphyrin functionalized graphene field effect transistor biosensor, the catalytic performance of graphene with good conductivity and specific porphyrin molecules on nitric oxide, the sensor the realization of ultra sensitive and specific detection of nitric oxide. The experiment proves that the biosensor exhibits excellent electrical conductivity and high Catalytic performance, nitric oxide can real-time monitoring of 1pM to 100nM range, the detection limit can be as low as 1pM, but also achieved at the single cell level ultra sensitive and high level of real-time monitoring of human umbilical vein endothelial cell specific release of nitric oxide. Due to its miniaturization, ultra sensitive and fast response characteristics of the biosensor greatly the potential application in biological and clinical diagnosis.

【學(xué)位授予單位】：湖北中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TN386;TP212.3

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