基于介电泳的细胞介电参数测试芯片机理的数值分析

设计一种集成四极式微电极组和螺旋微电极组的介电泳(Dielectrophoresis, DEP)细胞介电参数测试芯片,在芯片上集成了细胞的常规介电泳(Conventional DEP, CDEP)、行波介电泳(Travelling-wave DEP, TWDEP)和电旋转介电泳(Electrorotation, EROT)测试功能。运用相量法建立交流电场中细胞所受的常规介电泳力、行波介电泳力和介电泳旋转转矩的计算模型,推导出细胞水平速度和自旋角速度的表达式,并对本芯片的CDEP因子、TWDEP因子及EROT因子的大小和方向及细胞运动速度进行数值模拟并结合文献中的试验数据进行对比论证。结果表明：在高度大于20 m的区域,细胞沿螺旋电极径向的行进速度与细胞有效极化率的实部和虚部都相关,而细胞悬浮高度仅与有效极化率的实部相关;另一方面,电旋转最佳测试区域为近似正方形区域且每条边的中点均处于邻近的电极顶尖与电极腔中心连线的中点之上,此区域内的介电泳转矩变化量小于5%且介电泳力最小,利于电旋转测量。因此,本测试芯片能够通过多种介电泳模式更为全面、准确地获取细胞的介电特性信息。     

惯性力和介电泳力耦合的细胞分选仿真设计研究

针对目前微流控芯片单物理场分选精度低、通量小等劣势,提出了多物理场耦合的分选方式.多物理场粒子分选方案是采用惯性力与介电泳力耦合的方式,通过理论计算分析了惯性力与介电泳力的工作原理,并利用COMSOL对惯性力粒子速度变化、介电泳电场进行了仿真分析,对结构进行了优化,确定了芯片尺寸:整个通道由2部分组成,通道入口处利用惯...     

一种细胞电融合芯片的研制

描述了一种新的细胞电融合芯片的研究过程,分析芯片内微电极结构对电场大小、分布的影响.在前期细胞电融合芯片微电极研究的基础上,提出了更加适合于细胞电融合的电极阵列结构模型--梳状交叉微电极阵列,通过仿真计算和实际测试证明了该模型的有效性;通过对材料的优缺点进行分析,选择合适的芯片材料;硅基材料上的微电极及芯片加工工艺研究;微电极的抗氧化与抗腐蚀保护技术研究.最后,在新研制的细胞电融合芯片原型上进行了初步实验,取得了较好的实验效果.     

介电泳辅助的药物洗脱支架载药机理

针对微探针型药物洗脱支架局部载药的特殊要求,提出基于介电泳自组织的载药新方法。首先根据介电泳装配原理,建立介电泳辅助的药物洗脱支架载药模型,并结合量纲一介电泳力及粒子运动控制方程,对解空间内介电泳力分布及粒子运动轨迹进行有限元模拟,讨论粒子相对电极位置及布朗运动对其装配效率的影响,在理论上证明基于介电泳装配的载药方案完全可行。最后在数值模拟的基础上,通过支架表面尖端载药测试平台,对理论预测进行了试验验证。试验结果表明：在去离子水中,当电压峰—峰值为10 V,频率处于0.1～1.0 MHz范围内时,直径4.4 mm的乳胶颗粒在正介电泳力作用下向尖端聚集,且当频率约为500 kHz时,其粒子运动速度达到最大。     

基于介电电泳的粒子分离微流控芯片的研究

文中制备了一种用于粒子分离的介电电泳微流控芯片,利用粒子介电性质不同实现粒子批量、高效分离。采用MEMS工艺,由光刻有电极的ITO玻璃基底和PDMS微通道制备而成。在此基础上,测定了当缓冲溶液的电导率为1μS/cm、交流信号电压为10 V时聚苯乙烯小球和酵母菌的正负介电泳响应,确定了两种微粒的分离条件:酵母菌细胞在1~50 k Hz时表现负介电泳响应,50 k Hz~5 MHz时表现正介电泳响应,50 k Hz为交叉频率;聚苯乙烯小球在1 k Hz~5 MHz始终表现负介电泳响应。选取10 V、5 MHz交流电压信号作为分离条件,对直径均为5μm的聚苯乙烯小球和酵母菌进行了分离,分离效率达到92.4%。     

集成阻抗识别的介电泳芯片设计及其关键工艺

以微粒的阻抗识别为目的,设计出一种三维网格阵列微电极传感器.传感器将介电泳分离与阻抗识别集成在同一芯片上.首先通过介电泳将微粒中的一种分离至测试区域,再通过阻抗信号描述微粒特性.针对芯片关键区域,提出了调节SU8曝光时间以提高阻抗测试有效信号的工艺解决方案.实验表明,SU8最佳曝光时间为70 s.对芯片性能的相关实验表明,芯片可对不同直径的乳胶微粒进行介电泳分离与阻抗识别.     

实时可重构的共面型光诱导介电泳微操纵平台

归纳与总结了各类单细胞微操作技术的最新研究进展,提出一种新的实时可重构的共面型光诱导操控平台,完成了光诱导介电泳芯片的设计、加工与操控平台的搭建.在通过采用机器视觉及模式识别技术实现微粒自动识别与实时追踪的基础上,通过视觉反馈控制在光电导层上生成动态可重构的光模式虚拟电极阵列,形成特定的空间势能形貌,并基于介电泳效应实现微粒的可编程操作.最后,基于多物理场耦合模型及数值模拟技术,定量分析了新式光诱导介电泳芯片的捕获及释放模式,并基于临界最大流速Q这一品质因数,对比分析了共面型与经典式光诱导介电泳捕获模型的尺寸选择性与槽道高度相关性.研究结果表明,共面型光诱导介电泳模型相对于经典光电子镊而言,在单细胞排布方面更具有优势--更高的捕获强度、稳定性、选择性以及分选效率.     

半月形电极微液滴驱动的微流控芯片

针对目前数字微流控芯片驱动电压比较高的问题,本文对比传统的驱动电极结构,研制了一种可以降低驱动电压的半月形驱动电极数字微流控芯片.首先,基于介电湿润原理,分析微液滴所受介电湿润力和微液滴接触圆上有效三相接触线所对应弦长的关系.接着,对比分析了传统的方形、叉齿形驱动电极与提出的半月形驱动电极上液滴有效三相接触线所形成的弦长大小;分析得出3种驱动电极结构中半月形驱动电极所形成的有效弦长最大,从而表明半月形驱动电极的数字微流控芯片上介电驱动力最大.最后,利用设计制作的3种驱动电极介电湿润芯片分别实验验证驱动液滴的效果.结果表明,所研制的半月形驱动电极数字微流控芯片的最小驱动电压分别比方形和叉齿形驱动电极芯片降低了约37％和67％.另外,当有效驱动电压为60 V时,半月形驱动电极芯片上2μL去离子水微液滴的速度约为10 cm/s,分别是方形与叉齿形驱动电极芯片上液滴速度的3倍和2倍.得到的实验数据证明了半月形驱动电极数字微流控芯片实现了降低芯片驱动电压的目的.     

电连接器接触件结构分析与结构参数优化设计

为研究接触件结构参数对分离力和塑性变形的综合影响,对电连接器接触件进行了结构分析,提出一种接触件收口的尺寸计算方法,建立接触件分离力与结构参数的理论公式.计算实例应用均匀拉丁方试验设计方法对关键结构参数进行采样,以接触件收口后的塑性应变和分离力为优化目标函数,结合响应面近似建模技术和遗传算法,获得了最优的关键结构参数组合.通过对接触件优化前后的实验对比论证,验证了优化结果的正确性和优化方法的有效性.     

介电弹性体隔振平台设计与多物理场耦合仿真分析

针对航空航天等领域轻质结构的隔振问题,考虑到隔振性能和可靠性的要求,设计了一种介电弹性体半主动隔振平台,并结合自由能理论建立了该隔振系统的机电耦合动力学参激振动微分方程。将介电弹性体材料力电耦合模拟材料子程序(UMAT)嵌入Abaqus非线性有限元分析软件,进行多物理场耦合仿真分析,分析介电弹性体半主动隔振系统共振频率、模态振型与频响函数,从而得到其隔振特性与适用范围。     

一种压阻式微压力传感器

微压力传感器是微机电领域最早开始研究并且实用化的微器件之一，它结构简单、用途广。基于压阻效应、惠斯顿电桥等相关知识设计了一种压阻式微压力传感器。为增大灵敏度，设计了一种折弯形的压敏电阻。基于一些相关的微加工工艺制定了制作这种微传感器的工艺流程并且制作成功了传感芯片。设计了一个处理电路去获得此传感器的输出信号，它由两级放大电路和两级巴特沃斯低通滤波电路组成。最后利用这个测试系统检测出了随压力变化而发生变化的微电压信号。     

Theoretical investigation on nanoparticle concentrations in optoelectrofluidic chip based on diffusion,convection, and migration

A numerical model and simulation relative to an optoelectrofluidic chip has been presented in this article. Both dielectrophoretic and electroosmotic force attracting the nano-sized particles could be studied by the diffusion, convection, and migration equations. For the nano-sized particles, the protein with radius 3.6?nm is considered as the objective particle. The electroosmosis dependent upon applied frequency is calculated, which range 10²?Hz from 10⁸?Hz, and provides the much stronger force to enrich proteins than dielectrophoresis (DEP). Meanwhile, the induced light pattern size significantly affecting the concentration distribution is simulated. In this end, the concentration curve has verified that the optoelectrofluidic chip can be capable of manipulating and assembling the suspended submicron particles.     

敞开式离子源和微型气泵集成的微型针-柱放电结构芯片设计与实验分析

采用LIGA(Lithographie,Galvanoformung,Abformung)工艺设计加工了一种微型针-柱放电结构芯片,可实现敞开式离子源和微型气泵的系统集成。此微型放电结构由采用电镀铜加工而成的针电极、上圆柱电极和下圆柱电极组成,针-柱间距有1mm、2mm两种规格。在室温、大气压环境、无外界通入气流的条件下,通过在针-柱电极上加载负直流高压,可产生稳定的气体放电。利用风速计testo 405-V1测量芯片气体放电产生的离子风风速,结果表明针-柱间距为2mm时产生的离子风流速最大,可达0.79m/s。对针-柱间距2mm规格的微型芯片进行乙酸进样电离实验,当针-柱之间产生稳定的电晕放电后,可发现位于芯片出口处已被去离子水润湿的PH试纸变红。此时利用微弱电流检测系统采集电离产生的离子,当放电电压为-3300V时,通过微弱电流采集系统检测到的电流信号可达120pA。采用LTQ XL离子阱质谱仪,对芯片电离丙酮、无水乙醇和乙酸乙酯的离子产物进行检测,所得到的主要物质为质子化单体离子和二聚物离子。放电产生离子风风速以及乙酸的进样和电离实验表明,基于LIGA的微型针-柱结构芯片可实现大气压环境下敞开式离子源和微型气泵的双重功能。     

Curcumin disturbed cell‐cycle distribution of HepG2 cells via cytoskeletal arrangement

Due to its extensive antitumor activity, curcumin has been focused on by more researchers. But, its antiproliferative mechanisms are still unknown. Here we studied the antiproliferative activity of curcumin in human liver cancer HepG2 cells. In order to analyze the cytotoxic activity and anticancer mechanisms of curcumin, we carried out cytotoxicity tests using 3‐[4,5‐dimethyl‐2‐thiazolyl]‐2,5 diphenyltetrazolium bromide (MTT) assay. The HepG2 cell cycle distribution and the expression of tubulin were detected by flow cytometry. Alterations in morphological and cytoskeletal properties of HepG2 cells were investigated using atomic force microscopy (AFM). Simultaneously, the effects of curcumin on the growth and proliferation of HepG2 cells were also assayed by MTT method. Cells were incubated with different doses of curcumin (0–80 μmol/l) for 24 h, the cell viability decreased from 91.10 ± 3.2% to 10.84 ± 4.0%, and the 50% inhibiting concentration (IC₅₀) was 23.15 ± 0.37 μmol/l. Moreover, flow cytometry quantitatively detected that curcumin treatment resulted in a dose‐dependent accumulation of HepG2 cells in G2/M phase with concomitant losses from G0/G1 phase, so curcumin caused cell‐cycle arrest at G2/M phase. Furthermore, we discovered that curcumin was able to upregulate the expression of tubulin in HepG2 cells. In addition, AFM analysis including cell‐membrane structure and cytoskeleton networks is helpful to explain the relationship between the changes of cells and external pharmacologic stimulation. SCANNING 35: 253‐260, 2013. © 2012 Wiley Periodicals, Inc.     

Planar patch-clamp force microscopy on living cells

Here we report a new combination of the patch-clamp technique with the atomic force microscope (AFM). A planar patch-clamp chip microstructured from borosilicate glass was used as a support for mechanical probing of living cells. The setup not only allows for immobilizing even a non-adherent cell for measurements of its mechanical properties, but also for simultaneously measuring the electrophysiological properties of a single cell. As a proof of principle experiment we measured the voltage-induced membrane movement of HEK293 and Jurkat cells in the whole-cell voltage clamp configuration. The results of these measurements are in good agreement with previous studies. By using the planar patch-clamp chip for immobilization, the AFM not only can image non-adhering cells, but also gets easily access to an electrophysiologically controlled cellular probe at low vibrational noise.     

Effects of chip geometries on dielectrophoresis and electrorotation investigation

The electric fields employed for such work are generated using chips, such as planar linear interdigitated arrays or two parallel arrays. However, chip geometries usually affect the investigation of dielectrophoresis （DEP） and electrorotation （ER） significantly, and even may misdirect the theoretical prediction. In order to understand the electrodes geometries effect and provide a suitable range of parameters, three-dimensional simulations for the DEP and ER characterizations on the quadrupolar hyperbolical electrodes are carried out. Influences of the electrodes gaps, cell height, work region, energized voltage and frequencies for the DEP and ER manipulations are analyzed, and the analysis results show that the gaps of the electrodes and the cell height have enormous effects, but the work region is not so important. Moreover, depending on the theoretical analysis, ER experiments for polystyrene microspheres with the diameter of 20 ~m are carried out on two kinds of chips. The experimental results show that the microspheres rotate in the counter-field direction and the maximum rotation speed appears in the megahertz range. In addition, the experimental results are compared with the simulation results, showing that the three-dimensional simulations considering the chip geometries are more accurate than the two-dimensional predictions. This paper provides a new understanding for the theoretical predictions of DEP and ER manipulations, which decreases the difference of the theoretical and experimental results significantly, and will be significant for the lab chip research.     

基于透射式荧光型光纤生物传感器的结构设计

本文设计的基于透射式荧光型光纤生物传感器的样品反应池,采用光学技术和微流控技术、介电电泳及微/纳机械加工技术等,克服了现有同类仪器相对缺乏且多为断点测量的不足。该设计采用的微流控通道和温控系统保证生化样品在模拟生理条件下进行检测,真实地反映生物组分的特性;介电电泳将探针分子聚集到生物生物传感器的反应活性表面,保证靶标分子能探针分子的杂交,有利于微、痕量组分的富集;镜面的锥形腔体结构为聚光装置,然后将光信号耦合到光导纤维,导入光学型光纤生物传感器的检测器加工处理。该设计能缩短响应时间,提高生物传感器的检测灵敏度。     

High‐throughput continuous flow femtosecond laser‐assisted cell optoporation and transfection

We present a femtosecond‐laser based nanoprocessing system for transient optical cell membrane poration to allow the introduction of foreign molecules into the interior of a cell with very high throughput. In the setup, cells flow through a micro‐flow tube for spatial confinement and are simultaneously targeted by fs laser radiation. Beam‐shaping generates a focal geometry along a line which is scanned across the micro‐flow cell to increase the number of reachable cells. Successful cell membrane poration was observed indirectly by cell transfection even with cell‐light interaction times in the millisecond range. The system was characterized by experiments with Chinese hamster ovary cells regarding cell viability, the uptake of extrinsic molecules and cell transfection efficiency. The continuous flow of cells enables a tremendous increase of cell throughput compared to previous nonflow approaches by treating millions of cells, although with only limited efficiency. The setup opens the possibility to realize a completely automated high‐throughput laser‐assisted cell‐poration system which could be integrated in lab‐on‐a‐chip devices. Microsc. Res. Tech. 77:974–979, 2014. © 2014 Wiley Periodicals, Inc.