本文選題：味覺(jué) + 細(xì)胞芯片　； 參考：《浙江大學(xué)》2006年碩士論文

【摘要】：味覺(jué)作為人類(lèi)基本感覺(jué)之一,其研究卻遠(yuǎn)遠(yuǎn)落后于視覺(jué)、聽(tīng)覺(jué)等。不僅如此,目前的味覺(jué)電生理記錄,仍主要依賴(lài)于以膜片鉗為代表的細(xì)胞膜微電極穿刺或鉗制技術(shù)。該類(lèi)方法因其對(duì)細(xì)胞的穿刺損害作用,難以實(shí)現(xiàn)長(zhǎng)時(shí)程測(cè)量,同時(shí)也難以實(shí)現(xiàn)對(duì)細(xì)胞網(wǎng)絡(luò)的多位點(diǎn)同時(shí)測(cè)量。因此,研究者們開(kāi)發(fā)出基于微電極陣列(Microelectrode Array,MEA)、場(chǎng)效應(yīng)管(Field Effect Transistor,FET)陣列以及光尋址電位傳感器(Light Addressable Potentiometric Sensor,LAPS)的細(xì)胞芯片,其實(shí)質(zhì)就是在芯片表面培養(yǎng)可興奮性細(xì)胞如神經(jīng)元等,細(xì)胞通過(guò)一層薄的電解液同芯片的電極或柵極相耦合,從而芯片可以實(shí)時(shí)監(jiān)測(cè)藥物或者電刺激下細(xì)胞的電生理變化。細(xì)胞芯片可對(duì)多個(gè)細(xì)胞同時(shí)進(jìn)行長(zhǎng)期、無(wú)損的胞外電信號(hào)測(cè)量,已在細(xì)胞電生理研究、藥物篩選以及環(huán)境監(jiān)測(cè)等生物醫(yī)學(xué)領(lǐng)域得到了廣泛應(yīng)用。 本文結(jié)合我們實(shí)驗(yàn)室多年來(lái)在人工味覺(jué)和細(xì)胞芯片方面的研究,提出了基于LAPS的味覺(jué)細(xì)胞芯片。味覺(jué)受體細(xì)胞具有神經(jīng)細(xì)胞特性,也屬于可興奮性細(xì)胞,將其培養(yǎng)在LAPS芯片表面,味覺(jué)細(xì)胞電位的改變即可通過(guò)檢測(cè)LAPS的光生電流得以測(cè)量。與MEA和FET陣列芯片相比,LAPS芯片不僅制各工藝簡(jiǎn)單,而且其光源可移動(dòng)尋址,方便對(duì)芯片表面隨機(jī)培養(yǎng)的細(xì)胞進(jìn)行跟蹤定位,克服了MEA和FET陣列芯片固定檢測(cè)位點(diǎn)幾何特性的限制。 采用微機(jī)械加工技術(shù),我們制備了LAPS味覺(jué)細(xì)胞芯片,并用多聚鳥(niǎo)氨酸和層粘素混合的磷酸緩沖液處理以增強(qiáng)細(xì)胞在芯片表面的黏附性。采用微吸管吸取技術(shù),我們體外分離出大鼠的味蕾,將其培養(yǎng)在LAPS芯片上。選取去甲腎上腺素和河豚毒素作為刺激藥物,作用于芯片上已貼壁生長(zhǎng)的味覺(jué)細(xì)胞,驗(yàn)證了LAPS芯片檢測(cè)味覺(jué)細(xì)胞動(dòng)作電位變化的可行性。然后,選擇四種基本味質(zhì)(NaCl,HCl,蔗糖,MgSO_4)作用于味覺(jué)細(xì)胞,檢測(cè)LAPS的響應(yīng)。采用LabVIEW采集數(shù)據(jù)、Matlab進(jìn)行時(shí)域和頻域分析,我們定性分析了不同味質(zhì)作用于味覺(jué)細(xì)胞后胞外電信號(hào)的變化,證實(shí)了大鼠菌狀乳頭的味覺(jué)細(xì)胞對(duì)于咸味和酸味的敏感性,實(shí)現(xiàn)了細(xì)胞芯片在味覺(jué)傳導(dǎo)機(jī)理研究中的應(yīng)用。
[Abstract]:Taste sense as one of the basic human senses, its research lags far behind the visual, auditory, and so on. Moreover, the current taste electrophysiological records still rely mainly on the membrane microelectrode puncture or clamp technology represented by patch clamp. Because of its damage to cell puncture, this method is difficult to realize long-term measurement and multi-site simultaneous measurement of cell network. So researchers have developed a cell chip based on microelectrode arrays, Field Effect transistorization arrays and Light Addressable Potentiometric sensors (LAPs), which essentially grow excitable cells, such as neurons, on the surface of the chip. The cell is coupled with the electrode or grid of the chip through a thin layer of electrolyte, so that the chip can monitor the electrophysiological changes of the cell in real time under drug or electrical stimulation. Cell chips can be used to measure long-term, non-destructive extracellular electrical signals on many cells simultaneously, and have been widely used in cell electrophysiology research, drug screening and environmental monitoring and other biomedical fields. In this paper, a taste cell chip based on LAPS is proposed based on the research of artificial taste and cell chip in our laboratory for many years. Taste receptor cells are neuronal and excitable cells. When cultured on LAPS microarray, the changes of taste cell potential can be measured by detecting photogenerated currents of LAPS. Compared with MEA and FET chips, laps chip is not only easy to manufacture, but also its light source can be moved and addressable, which makes it easy to track and locate the cells cultured randomly on the surface of the chip. It overcomes the limitation of geometric characteristics of fixed detection sites in MEA and FET array chips. A LAPS taste cell chip was prepared by micromachining and was treated with polyornithine and laminin phosphate buffer to enhance cell adhesion on the chip surface. The taste buds of rats were isolated by micropipette extraction and cultured on LAPS microarray. Norepinephrine and tetrodotoxin were selected as stimulators to act on the taste cells grown on the chip. The feasibility of detecting the action potential of taste cells by LAPS chip was verified. Then, four basic flavors, NaCl-HCl, Sucrose Mg-SO4) were selected to act on the taste cells to detect the response of LAPS. Time domain and frequency domain analysis were carried out with LabVIEW data. We qualitatively analyzed the changes of extracellular electrical signals after different taste substances acted on taste cells, and confirmed the sensitivity of taste cells of rat papilla to salty and sour taste. The application of cell chip in the study of taste conduction mechanism is realized.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類(lèi)號(hào)】：R33

【引證文獻(xiàn)】

相關(guān)期刊論文 前3條

1 陳培華;劉曉東;張威;周俊;王平;楊巍;羅建紅;;味覺(jué)感受細(xì)胞離子通道和動(dòng)作電位的建模與仿真[J];中國(guó)科學(xué)(C輯:生命科學(xué));2009年02期

2 呂曉迎;;生物電子學(xué)領(lǐng)域中的生物材料與生物相容性研究[J];中國(guó)材料進(jìn)展;2010年12期

3 陳培華;張威;周俊;王平;肖麗丹;楊莫;;膜片鉗芯片技術(shù)及其在細(xì)胞電生理分析中的研究進(jìn)展[J];自然科學(xué)進(jìn)展;2007年12期

相關(guān)博士學(xué)位論文 前1條

1 吳春生;仿生嗅覺(jué)細(xì)胞及受體傳感器的研究[D];浙江大學(xué);2009年

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本文編號(hào)：1909627