本文選題：細(xì)胞電融合　切入點(diǎn)：微流控芯片　出處：《重慶大學(xué)》2015年碩士論文

【摘要】：細(xì)胞融合作為細(xì)胞工程的主要技術(shù)手段之一,在胚胎學(xué)、醫(yī)藥學(xué)、免疫學(xué)、遺傳學(xué)、食品營(yíng)養(yǎng)學(xué)等學(xué)科有重要應(yīng)用。作為細(xì)胞融合技術(shù)的主流手段之一,細(xì)胞電融合具有對(duì)細(xì)胞損傷小、無(wú)殘存毒性、融合率高、誤差小、便于研究者操控和觀察等優(yōu)點(diǎn),已被廣泛用于細(xì)胞融合領(lǐng)域。細(xì)胞電融合操作的設(shè)備是細(xì)胞融合裝置,國(guó)外細(xì)胞融合裝置雖能顯著提升細(xì)胞融合率,但其價(jià)格昂貴,配套融合池尺寸(毫米級(jí))過(guò)大限制了科研人員對(duì)細(xì)胞的精密操控,同時(shí)提升了功能電路工作電壓,影響融合后細(xì)胞的存活率;國(guó)內(nèi)細(xì)胞融合裝置雖然作用細(xì)胞種類多,但與國(guó)外細(xì)胞融合裝置相比,融合效率較低下,且功能電路工作不穩(wěn)定。針對(duì)上述事實(shí),本設(shè)計(jì)結(jié)合課題組進(jìn)行的微流控細(xì)胞電融合芯片研究,在前期研究初步實(shí)現(xiàn)的基于微電極芯片(尺寸微米級(jí))的細(xì)胞融合裝置的基礎(chǔ)上,提出了基于功能電路劃分的系統(tǒng)設(shè)計(jì),并重點(diǎn)進(jìn)行了功能電路的設(shè)計(jì)研究。該功能電路有提高細(xì)胞融合率、減少實(shí)驗(yàn)成本、提升實(shí)驗(yàn)操作的效率等優(yōu)點(diǎn),系統(tǒng)功能電路主要包括排隊(duì)模塊、擊穿模塊和其他模塊,其中,細(xì)胞排隊(duì)模塊由DDS信號(hào)發(fā)生器單元、信號(hào)濾波單元、幅度控制單元、放大電路單元組成;細(xì)胞擊穿模塊方面由TTL脈沖發(fā)生單元、MOS管驅(qū)動(dòng)單元、MOS管開(kāi)關(guān)單元、RC充放電單元組成;其他模塊由信號(hào)切換模塊、電源模塊、通訊模塊和控制模塊組成。本設(shè)計(jì)采用功能模塊化的設(shè)計(jì)思路,使功能電路模塊化,并測(cè)試了各模塊的信號(hào)參數(shù),結(jié)果表明參數(shù)滿足設(shè)計(jì)需求;利用C粉(碳粉)和PDMS復(fù)合電極開(kāi)展了功能電路的負(fù)載測(cè)試,結(jié)果表明信號(hào)波形完整,各模塊帶負(fù)載能力良好;借用課題組培養(yǎng)的乳腺癌細(xì)胞,結(jié)合排隊(duì)模塊分別在C粉與PDMS復(fù)合電極、銀針電極、銀粉與PDMS復(fù)合電極上做了初步的細(xì)胞排隊(duì)驗(yàn)證實(shí)驗(yàn),實(shí)驗(yàn)結(jié)果表明乳腺癌細(xì)胞在本設(shè)計(jì)功能電路作用下能夠在銀粉和PDMS復(fù)合電極上發(fā)生有效聚集,形成珍珠串,并且具有較高的細(xì)胞排隊(duì)率。
[Abstract]:As one of the main techniques in cell engineering, cell fusion plays an important role in embryology, medicine, immunology, genetics, food nutrition and so on.As one of the main methods of cell fusion, electrofusion has been widely used in the field of cell fusion because of its advantages of little damage to cells, no residual toxicity, high fusion rate, low error and easy to be manipulated and observed by researchers.Cell fusion is operated by cell fusion devices. Although foreign cell fusion devices can significantly improve cell fusion rates, they are expensive, and the size of matching fusion pools (millimeters) greatly limits the precise manipulation of cells by researchers.At the same time, the working voltage of the functional circuit is increased, and the survival rate of the fused cells is affected. Although there are many kinds of cells acting on the cells in our country, the fusion efficiency is lower and the functional circuit is unstable compared with the foreign cell fusion devices.In view of the above facts, this design combined with the research of microfluidic cell electrofusion chip carried out by our research group, on the basis of the preliminary implementation of the cell fusion device based on microelectrode chip (size and micron scale),The system design based on functional circuit partition is put forward, and the design of functional circuit is emphasized.The functional circuit has the advantages of improving the cell fusion rate, reducing the experimental cost and improving the efficiency of the experimental operation. The functional circuit of the system mainly includes queue module, breakdown module and other modules, among which,The cell queue module is composed of DDS signal generator unit, signal filter unit, amplitude control unit and amplification circuit unit, and the cell breakdown module is composed of TTL pulse generator unit, MOS tube driving unit, MOS tube switch unit and RC charging and discharging unit.Other modules are composed of signal switching module, power supply module, communication module and control module.The design adopts the idea of functional modularization, makes the functional circuit modularized, and tests the signal parameters of each module, the results show that the parameters meet the design requirements;The load test of functional circuit was carried out by using C powder (carbon powder) and PDMS composite electrode. The results show that the signal waveform is complete, and each module has good load capacity.Combined with queuing module, the cell queuing verification experiments were carried out on C powder and PDMS composite electrode, silver needle electrode, silver powder and PDMS composite electrode, respectively.The experimental results show that breast cancer cells can effectively aggregate on silver powder and PDMS composite electrode under the action of the designed functional circuit, form pearl string, and have high cell queuing rate.
【學(xué)位授予單位】：重慶大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：Q813.2;TN492

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