雙乳液滴內(nèi)核可控包裹與融合機(jī)制及實(shí)驗(yàn)研究
發(fā)布時(shí)間：2019-04-03 08:50
【摘要】：液滴微流控作為一種新的流體控制技術(shù)可以在液滴內(nèi)實(shí)現(xiàn)納升級(jí)別的微反應(yīng),已然成為微尺度上生物和化學(xué)反應(yīng)及其過(guò)程檢測(cè)的有效手段。液滴作為微反應(yīng)器,能夠有效控制擴(kuò)散、加速混合、提高檢測(cè)靈敏度,其應(yīng)用范圍涉及到單細(xì)胞分析、藥物篩選、個(gè)人洗護(hù)等多學(xué)科交叉領(lǐng)域。然而,單乳液滴在壁面潤(rùn)濕、界面擾動(dòng)等外界因素作用下易產(chǎn)生樣品泄露,造成交叉污染,且不易進(jìn)行多組分液滴操控。雙乳液滴獨(dú)特的核-殼結(jié)構(gòu)能夠有效保護(hù)內(nèi)核液滴中的反應(yīng)樣品及反應(yīng)產(chǎn)物,同時(shí)避免有害反應(yīng)產(chǎn)物對(duì)外界環(huán)境的污染,且多核雙乳液滴為多組分液滴的精準(zhǔn)操控提供了重要載體。液滴微流控芯片是一個(gè)研究平臺(tái),其目的是為各領(lǐng)域的應(yīng)用提供方法。但是,微尺度下的反應(yīng)具有高度的多樣化,單一的液滴微流控系統(tǒng)很難滿足多樣化的微反應(yīng)需求。因此,需要多種微液滴形式及操控方法實(shí)現(xiàn)不同條件下的微反應(yīng)器功能,從而去滿足多樣化的微反應(yīng)需求。本文針對(duì)上述問(wèn)題展開。從微納尺度流體流動(dòng)角度出發(fā),根據(jù)微通道內(nèi)兩相流的基本理論,分析了液滴的形成及水包油包水(Water-in-Oil-in-Water,W/O/W)型雙核雙乳液滴內(nèi)核電融合機(jī)理。建立了基于共軸流法的液滴生成仿真模型,對(duì)影響液滴生成大小及頻率的關(guān)鍵因素進(jìn)行了模擬分析,為液滴生成玻璃微流控芯片的設(shè)計(jì)提供支撐;建立了交流電場(chǎng)中W/O/W雙核雙乳液滴的受力及仿真二維模型,對(duì)內(nèi)核液滴在極化作用下的電場(chǎng)力和流體動(dòng)力進(jìn)行分析,得出電場(chǎng)頻率、電壓幅值、內(nèi)核電導(dǎo)率與內(nèi)核液滴電融合之間的影響關(guān)系,為后續(xù)實(shí)驗(yàn)研究奠定理論基礎(chǔ)。從簡(jiǎn)單實(shí)用角度考慮,設(shè)計(jì)了一種玻璃毛細(xì)管微流控芯片系統(tǒng),在不需要停止實(shí)驗(yàn)和更換溶液的情況下,實(shí)現(xiàn)了W/O/W雙乳液滴中兩種不同試劑的“一步”定比例包裹,在大量節(jié)省試劑的條件下可以完成多工況的納升級(jí)微反應(yīng);根據(jù)質(zhì)量守恒方程,推導(dǎo)了雙乳液滴大小及殼厚與各相流體流速的關(guān)系式,作為實(shí)驗(yàn)驗(yàn)證,合成了具有不同大小,不同殼厚,且具有高度均勻性的雙乳液滴;通過(guò)對(duì)雙乳液滴的PDMS(polydimethylsiloxane)外殼進(jìn)行恒溫固化,成功將雙乳液滴轉(zhuǎn)換成了具有一定機(jī)械強(qiáng)度的微囊泡結(jié)構(gòu),并對(duì)其進(jìn)行了長(zhǎng)達(dá)7個(gè)月的時(shí)間耐久性檢測(cè),確保其內(nèi)部包裹物可以長(zhǎng)時(shí)間儲(chǔ)存和觀察;最后,利用酶催化反應(yīng)和中和反應(yīng)驗(yàn)證了此方法在多工況微反應(yīng)器中應(yīng)用的可行性。從靈活可控角度考慮,設(shè)計(jì)并搭建了用于生成包含兩個(gè)不同內(nèi)核的W/O/W雙乳液滴實(shí)驗(yàn)平臺(tái),通過(guò)調(diào)節(jié)內(nèi)核液滴與外界介質(zhì)中的鹽濃度差異,研究了滲透壓作用下雙核雙乳液滴的膨脹和形變規(guī)律,包括兩內(nèi)核在不同滲透壓作用下隨時(shí)間等速膨脹后的融合和非等速膨脹后的單一內(nèi)核破裂。利用滲透壓誘導(dǎo)實(shí)現(xiàn)了雙乳液滴中分別含有Ca Cl2和Na2CO3溶液的兩個(gè)內(nèi)核液滴融合,進(jìn)而實(shí)現(xiàn)了其內(nèi)部溶液的混合及CaCO3生成反應(yīng),驗(yàn)證了滲透壓誘導(dǎo)內(nèi)核融合的雙核雙乳液滴作為納升級(jí)微反應(yīng)器的可行性。尤其是在沒(méi)有電能的惡劣條件下,僅僅通過(guò)鹽濃度的差異,就可以實(shí)現(xiàn)納升級(jí)微反應(yīng)。從即時(shí)操控角度考慮,利用交流電場(chǎng)的極化作用實(shí)現(xiàn)了連續(xù)流中W/O/W雙核雙乳液滴的內(nèi)核液滴即時(shí)融合,研究了連續(xù)流流速、電場(chǎng)頻率、電壓幅值、內(nèi)核溶液電導(dǎo)率等對(duì)大批量雙核雙乳液滴內(nèi)核液滴融合效率的影響關(guān)系。通過(guò)合理調(diào)節(jié)各參數(shù),連續(xù)流中W/O/W雙核雙乳液滴的內(nèi)核液滴融合效率可以達(dá)到95%以上,為大樣本需求的納升級(jí)微反應(yīng)提供技術(shù)支持。另外,通過(guò)酶催化的葡萄糖檢測(cè)反應(yīng)和水凝膠微粒合成及包裹酵母菌細(xì)胞的實(shí)驗(yàn),驗(yàn)證了內(nèi)核電融合的雙核雙乳液滴作為即時(shí)操控型納升級(jí)微反應(yīng)器的可行性。
[Abstract]:Microfluidic as a new kind of fluid control technology can realize the micro-reaction of nano-upgrading in the liquid drop, and has become an effective means of biological and chemical reaction in micro-scale and its process detection. The liquid drops act as micro-reactors, can effectively control the diffusion, accelerate the mixing, and improve the detection sensitivity, and the application range of the liquid drops relates to the cross-cutting fields of single-cell analysis, drug screening, personal care and the like. However, that single-emulsion drop is easy to generate sample leakage under the action of wall surface wetting, interface disturbance and other external factors, so that the cross-contamination is caused, and the multi-component liquid drop control is not easy to be carried out. The unique core-shell structure of the double-emulsion liquid drops can effectively protect the reaction sample and the reaction product in the inner core liquid drop, and meanwhile, the pollution of the harmful reaction product to the external environment is avoided, and the multi-core double-emulsion liquid drop is an important carrier for precise control of the multi-component liquid drop. The droplet micro-fluidic chip is a research platform, the purpose of which is to provide a method for the application of each field. However, the micro-scale reaction has a high degree of diversification, and a single droplet micro-flow control system is difficult to meet the diversified micro-reaction requirements. Therefore, the micro-reactor function under different conditions is realized by the form of a plurality of micro-droplets and the control method, so that the diversified micro-reaction requirements can be met. This paper is based on the above-mentioned problems. According to the basic theory of the two-phase flow in the micro-channel, the electric fusion mechanism of the water-in-water-in-Water (W/ O/ W) dual-core double-emulsion droplet is analyzed based on the basic theory of the two-phase flow in the micro-channel. The simulation model of the droplet generation based on the co-axial flow method is established, and the key factors that influence the size and frequency of the droplet generation are simulated and analyzed, and a support is provided for the design of the liquid drop generating glass micro-fluidic chip, and the force and the simulation two-dimensional model of the W/ O/ W dual-core double-emulsion liquid drop in the alternating-current electric field are established, The influence of the electric field frequency, the amplitude of the voltage, the electrical conductivity of the inner core and the electric fusion of the inner core is obtained, and the theoretical basis for the subsequent experimental research is obtained. from a simple and practical point of view, a glass capillary micro-fluidic chip system is designed, and the "one step" of two different reagents in the W/ O/ W double-emulsion liquid drop is realized under the condition of not stopping the experiment and changing the solution, According to the mass conservation equation, the relation between the size of the double-emulsion liquid drop and the shell thickness and the fluid flow rate of each phase is deduced, and the double-emulsion liquid drops are transformed into a micro-capsule structure with a certain mechanical strength by performing constant-temperature curing on the PDMS (polydimethylsiloxane) shell of the double-emulsion liquid drop, And finally, the feasibility of the application of the method in a multi-working condition micro-reactor is verified by using the enzyme catalysis reaction and the neutralization reaction. In this paper, we design and set up a W/ O/ W double-emulsion drop experiment platform for generating two different cores from a flexible and controllable angle. By adjusting the difference of the salt concentration between the inner core and the external medium, the expansion and deformation of the dual-core double-emulsion liquid drop under the influence of the osmotic pressure are studied. Comprises the fusion of the two cores under different osmotic pressure and the single core rupture after the non-constant-speed expansion. By using osmotic pressure induction, the two core liquid droplets containing the solution of Ca 2 and Na2CO3 in the double-emulsion liquid droplet were fused, and then the mixing of the internal solution and the reaction of CaCO3 formation were realized, and the feasibility of the dual-core double-emulsion liquid droplet fused by the osmotic pressure-inducing core as a nano-upgrading micro-reactor was verified. In particular, the nano-upgrading micro-reaction can be realized only by the difference of the salt concentration under the severe conditions of no electric energy. The instantaneous fusion of W/ O/ W dual-core double-emulsion droplets in continuous flow is realized by the polarization effect of the AC electric field from the point of view of the real-time operation. The continuous flow velocity, the frequency of the electric field and the amplitude of the voltage are studied. The effect of the electrical conductivity of the core solution on the droplet fusion efficiency of the large-scale dual-core double-emulsion liquid droplet. By reasonably adjusting each parameter, the fusion efficiency of the inner core of the W/ O/ W dual-core double-emulsion liquid drop in the continuous flow can reach more than 95%, and technical support is provided for the nano-upgrading micro-reaction of the large sample requirement. In addition, by enzyme-catalyzed glucose test and water-gel particle synthesis and the experiments of the yeast cells, the feasibility of the dual-core dual-emulsion liquid drop as an immediate control type micro-reactor was verified.
【學(xué)位授予單位】：哈爾濱工業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：O35
本文編號(hào)：2453082
[Abstract]:Microfluidic as a new kind of fluid control technology can realize the micro-reaction of nano-upgrading in the liquid drop, and has become an effective means of biological and chemical reaction in micro-scale and its process detection. The liquid drops act as micro-reactors, can effectively control the diffusion, accelerate the mixing, and improve the detection sensitivity, and the application range of the liquid drops relates to the cross-cutting fields of single-cell analysis, drug screening, personal care and the like. However, that single-emulsion drop is easy to generate sample leakage under the action of wall surface wetting, interface disturbance and other external factors, so that the cross-contamination is caused, and the multi-component liquid drop control is not easy to be carried out. The unique core-shell structure of the double-emulsion liquid drops can effectively protect the reaction sample and the reaction product in the inner core liquid drop, and meanwhile, the pollution of the harmful reaction product to the external environment is avoided, and the multi-core double-emulsion liquid drop is an important carrier for precise control of the multi-component liquid drop. The droplet micro-fluidic chip is a research platform, the purpose of which is to provide a method for the application of each field. However, the micro-scale reaction has a high degree of diversification, and a single droplet micro-flow control system is difficult to meet the diversified micro-reaction requirements. Therefore, the micro-reactor function under different conditions is realized by the form of a plurality of micro-droplets and the control method, so that the diversified micro-reaction requirements can be met. This paper is based on the above-mentioned problems. According to the basic theory of the two-phase flow in the micro-channel, the electric fusion mechanism of the water-in-water-in-Water (W/ O/ W) dual-core double-emulsion droplet is analyzed based on the basic theory of the two-phase flow in the micro-channel. The simulation model of the droplet generation based on the co-axial flow method is established, and the key factors that influence the size and frequency of the droplet generation are simulated and analyzed, and a support is provided for the design of the liquid drop generating glass micro-fluidic chip, and the force and the simulation two-dimensional model of the W/ O/ W dual-core double-emulsion liquid drop in the alternating-current electric field are established, The influence of the electric field frequency, the amplitude of the voltage, the electrical conductivity of the inner core and the electric fusion of the inner core is obtained, and the theoretical basis for the subsequent experimental research is obtained. from a simple and practical point of view, a glass capillary micro-fluidic chip system is designed, and the "one step" of two different reagents in the W/ O/ W double-emulsion liquid drop is realized under the condition of not stopping the experiment and changing the solution, According to the mass conservation equation, the relation between the size of the double-emulsion liquid drop and the shell thickness and the fluid flow rate of each phase is deduced, and the double-emulsion liquid drops are transformed into a micro-capsule structure with a certain mechanical strength by performing constant-temperature curing on the PDMS (polydimethylsiloxane) shell of the double-emulsion liquid drop, And finally, the feasibility of the application of the method in a multi-working condition micro-reactor is verified by using the enzyme catalysis reaction and the neutralization reaction. In this paper, we design and set up a W/ O/ W double-emulsion drop experiment platform for generating two different cores from a flexible and controllable angle. By adjusting the difference of the salt concentration between the inner core and the external medium, the expansion and deformation of the dual-core double-emulsion liquid drop under the influence of the osmotic pressure are studied. Comprises the fusion of the two cores under different osmotic pressure and the single core rupture after the non-constant-speed expansion. By using osmotic pressure induction, the two core liquid droplets containing the solution of Ca 2 and Na2CO3 in the double-emulsion liquid droplet were fused, and then the mixing of the internal solution and the reaction of CaCO3 formation were realized, and the feasibility of the dual-core double-emulsion liquid droplet fused by the osmotic pressure-inducing core as a nano-upgrading micro-reactor was verified. In particular, the nano-upgrading micro-reaction can be realized only by the difference of the salt concentration under the severe conditions of no electric energy. The instantaneous fusion of W/ O/ W dual-core double-emulsion droplets in continuous flow is realized by the polarization effect of the AC electric field from the point of view of the real-time operation. The continuous flow velocity, the frequency of the electric field and the amplitude of the voltage are studied. The effect of the electrical conductivity of the core solution on the droplet fusion efficiency of the large-scale dual-core double-emulsion liquid droplet. By reasonably adjusting each parameter, the fusion efficiency of the inner core of the W/ O/ W dual-core double-emulsion liquid drop in the continuous flow can reach more than 95%, and technical support is provided for the nano-upgrading micro-reaction of the large sample requirement. In addition, by enzyme-catalyzed glucose test and water-gel particle synthesis and the experiments of the yeast cells, the feasibility of the dual-core dual-emulsion liquid drop as an immediate control type micro-reactor was verified.
【學(xué)位授予單位】：哈爾濱工業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：O35
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1 侯立凱;雙乳液滴內(nèi)核可控包裹與融合機(jī)制及實(shí)驗(yàn)研究[D];哈爾濱工業(yè)大學(xué);2017年
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