基于RGD多肽掺杂聚吡咯-铟锡氧化物的仿生微电极构建及用于细胞生物学行为的电化学阻抗谱检测

构建一种基于RGD多肽分子掺杂聚吡咯膜修饰的铟锡氧化物微电极(PPy/RGD-ITO),并以此作为传感电极实现细胞生物学行为的电化学阻抗谱检测.采用光刻技术蚀刻感光干膜绝缘层制备ITO微电极;以含RGD模体的多肽分子作为吡咯电聚合唯一的掺杂阴离子,通过电化学共聚合方式在ITO微电极表面沉积PPy/RGD复合膜形成PPy/RGD-ITO微电极;原子力显微镜(AFM)、接触角测量仪和傅里叶变换红外光谱仪(FTIR)分别表征PPy/RGD复合膜的表面拓扑形貌、湿润性和组成成分;人肺癌细胞株A549铺展、粘附及增殖实验考察了PPy/RGD复合膜与细胞间的相互作用;以构建的PPy/RGD-ITO微电极作为传感电极,通过电化学阻抗谱技术对A549细胞粘附增殖行为及天然抗癌药物分子重楼皂苷I的细胞毒性进行了分析.结果显示,通过简单的电化学共聚合成功将RGD分子掺杂进PPy膜内,且PPy/RGD复合膜具有优异的表面物理性能;PPy基质膜内掺杂的RGD分子保留其生物活性,相比裸ITO电极和聚4-苯乙烯磺酸钠(PSS)掺杂的PPy膜,PPy/RGD复合膜能更好地促进A549细胞的铺展、粘附和增殖;由于PPy/RGD-ITO微电极表面A549细胞形态学变化可改变电极系统的阻抗谱特征,因此通过电化学阻抗谱技术可解析A549细胞粘附增殖行为学信息,同时可定量分析重楼皂苷I细胞毒性.因此,通过简单的电化学共聚合方法将生物活性RGD分子掺杂进PPy膜内制备出的PPy/RGD膜具有优良的生物相容性,可作为一种重要的仿生电极修饰材料用于构建电子系统和细胞生物学系统的耦合界面,未来可应用于细胞生物学行为及药物筛选研究.     

一种基于感光干膜-铟锡氧化物电极的简易细胞阻抗传感器实现细胞形态学和阻抗信息同时检测

加工一种基于感光干膜-铟锡氧化物DFP-ITO( Dry Film Photoresist-Indium Tin Oxide)电极的细胞阻抗生物传感器并实现细胞形态学和阻抗信息同时检测。35μm厚的感光干膜层压在ITO导电玻璃表面上作为绝缘层,通过照相制版技术在感光干膜绝缘层上蚀刻不同直径圆孔;以DFP-ITO作为工作电极,通过夹具和测量小池与Ag/AgCl参比电极、Pt丝对电极相连构成三电极阻抗测量系统;考察了不同直径DFP-ITO工作电极阻抗谱特征;通过细胞粘附实验及细胞毒性实验考察了感光干膜细胞生物相容性;通过光学显微镜和阻抗谱技术分别对接种在DFP-ITO电极上人肺癌细胞株A549粘附、增殖过程中的形
　　态学和阻抗信息进行检测和分析。研究结果发现不同直径DFP-ITO电极具有相似的阻抗特性;充分固化的感光干膜表面适宜A549细胞粘附且无明显的细胞毒性;基于DFP-ITO电极构建的细胞阻抗传感器能够通过光学显微镜获取A549细胞形态学数据,同时通过阻抗谱技术能够解析A549细胞粘附、增殖过程中的细胞质膜电容、细胞-细胞间隙电阻、细胞-ITO电极间隙电阻变化。本文发展了基于DEP-ITO电极的细胞阻抗传感器结构简单,可实现细胞形态学和阻抗信息的双通道获取,未来可用于细胞生理病理学行为和药物细胞毒性研究。     

微电极阵列细胞阻抗传感器研究

贴壁生长在微电极表面的细胞可引起贴壁电极界面阻抗的改变,从而可以获得细胞生理功能相关的生物信息.本研究采用微机械加工技术,在硅基底上设计了直径为20～50 μm的20通道金微电极阵列(micro-electrode array,MEA),用以构建能实时、连续、定量跟踪哺乳动物细胞形态和增殖分化改变的细胞阻抗测试传感器(electric cell-substrate impedance sens-ing,ECIS),用于细胞与电极间的阻抗测试研究.通过对培养在微电极表面24 h的成骨细胞Saos-2细胞系的阻抗谱测量发现,其阻抗值增加集中在中频102～10<'4>Hz之间,本结果符合细胞阻抗传感测量的理论模型分析.因此,该微电极阵列细胞阻抗传感器的研究,为进一步的细胞生理和药物分析等研究奠定了良好的基础.     

基于柔性衬底的人造视网膜生物微电极阵列研究

设计了一种用于刺激视网膜的生物微电极阵列,采用非硅MEMS技术,在聚酰亚胺柔性衬底上制备出了具有一定生物相容性的,电极数为20的生物刺激电极阵列,并成功实现了器件从基底的完整释放。实验中采用了二次曝光法,用金修饰了电极柱侧壁,从而提高了电极的生物相容性;采用PDMS作为柔性衬底与玻璃片的粘附层,使得器件的释放过程,简单且无毒无害,对器件也无损害。制造出的器件尺寸小,质量轻,可靠性高,机械柔性好。对微电极阵列进行了电学性能测试,在10~5000Hz频率范围,其阻抗为104~106欧姆,符合电刺激要求。     

快速检测大肠杆菌O157:H7的电化学阻抗免疫生物传感器

建立了一种采用电化学阻抗谱技术快速检测大肠杆菌O157∶H7的生物传感器,它是通过石英晶体金电极表面附着一层蛋白A膜来固定抗体的。该生物传感器采用了三电极系统-工作电极石英晶体金电极、Ag/AgCl/Cl-SAT参考电极和铂对电极。和以往的普通金电极不同,第一次采用了石英晶体金电极。试验结果说明抗体的固定以及大肠杆菌O157∶H7与抗体的结合都增加了石英晶体金电极表面的电子传递阻抗,在[Fe(CN)6]3-/4-氧化还原对存在的情况下,用电化学阻抗谱测量该阻抗。该免疫生物传感器的检测限是103cfu/mL,石英晶体金电极的电子传递阻抗变化值和大肠杆菌O157∶H7的浓度在一定范围内呈线性关系,检测时间少于10 min。     

微电子细胞传感器在抗肿瘤药物分析中的应用研究

研究采用微机械加工技术制备了生物电阻抗检测用的细胞传感器芯片。传感器各通道为50μm的微电极,以交错的方式间隔50μm排列。配以相应的药物微进样系统与控制系统后,将人肺腺癌细胞系A549细胞接种培养于芯片电极表面,研究了人参皂苷Rh2对细胞的作用。阻抗测试结果显示:人参皂苷Rh2具有良好的抑制肿瘤细胞生长作用,表明细胞微电子芯片为抗肿瘤药物的分析与筛选提供了一个良好微传感器测试平台。     

基于柔性衬底的三维生物刺激微电极阵列研究

设计了一种基于聚酰亚胺薄膜的三维生物刺激微电极阵列,用于植入式人造视网膜应用.采用非硅MEMS技术,在柔性衬底上制备出具有生物相容性和化学稳定性,电极高度为80 μm的生物刺激电极阵列,通过PDMS牺牲层实现器件从基底的完整释放.实验中器件以聚酰亚胺和PDMS封装,电极柱和焊盘均镀金,从而提高电极的生物相容性.采用三电极法对微电极进行了电化学性能测试,在10-1～105Hz频率范围内,其阻抗为1.5～0.3 kΩ.制造出的器件尺寸小,质量轻,可靠性高,机械柔性好,符合生物电刺激要求.     

聚吡咯掺杂苯磺酸钠葡萄糖生物传感器

采用电化学聚合技术,用掺杂苯磺酸钠的聚吡咯（PPy）导电薄膜修饰铅笔芯电极,在修饰电极表面吸附葡萄糖氧化酶制备了葡萄糖生物传感器.研究了苯磺酸钠掺杂对PPy薄膜形貌、葡萄糖传感器性能的影响.实验结果表明：掺杂苯磺酸钠能够改变PPy形貌、极大提高其导电性.优化条件下该生物传感器抗干扰能力强、稳定性好,响应电流和葡萄糖浓度在0～0.7 mmol/L范围内有良好的线性相关度（R=0.9976）,灵敏度为26.10 μA/mmol/L,平均响应时间约为6.5s,检测下限为47.2 μmol/L.     

检测大肠杆菌O157:H7的电化学阻抗谱生物传感器的研究

我们提出了用掺锡的三氧化二铟(ITO)作为工作电极,通过硅烷化固定化技术,将抗大肠杆菌O157:H7单克隆抗体固定在ITO电极表面,利用电化学阻抗谱技术来构建一种新型的免疫传感器.该新型的免疫传感器的检测限为 4×103CFU/mL,检测线性范围为4×103-4×106CFU/mL.实验研究表明,该传感器具有灵敏度较高,检测时间短,操作简单等优点,在临床医学和环境监测中具有应用价值.     

基于MONTE-CARLO模型的细胞传感器纳米颗粒表面处理分析

离体生物传感器的微型和集成化为细胞电生理研究提供了有力手段.当前微电极阵列表面纳米粒子处理的相关量化分析尚为欠缺.着重采用Monte-Carlo模型判断单层/多层纳米粒子膜与电极耦合的有效性,讨论异质的单一或不同粒径分散度(50～500 am)的纳米粒子分层贴附处理于电极上,从二维、三维模型角度分析纳米粒子表面处理的有...     

Electrochemical deposition of polypyrrole/graphene oxide composite on microelectrodes towards tuning the electrochemical properties of neural probes

By exploiting the electrostatic interaction between positively charged pyrrole cation radicals and negatively charged graphene oxide (GO) sheets, we prepared polypyrrole/graphene oxide (PPy/GO) composite films by a one-step electrochemical process. We studied the effects of the polymerization current density and the GO content in electrolyte on the formation of PPy/GO coatings onto platinum neural microelectrode sites. As compared with pure PPy film, PPy/GO coatings show a rougher surface feature with micrometer-scale bulges. The impedance of the PPy/GO coated Pt electrode is only about 10% of the bare Pt electrode at the biological relevant 1 kHz, while the charge capacity density is more than two orders of the magnitude of the bare Pt electrode. Moreover, the PPy/GO coated Pt electrodes show higher performance than the PPy coated electrodes for the application of neural probe.     

A novel bienzyme glucose biosensor based on three-layer Au-Fe3O4@SiO2 magnetic nanocomposite

The magnetic core-shell Au-Fe₃O₄@SiO₂ nanocomposite was prepared by layer-by-layer assembly technique and was used to fabricate a novel bienzyme glucose biosensor. Glucose oxidase (GOD) and horseradish peroxidase (HRP) were simply mixed with Au-Fe₃O₄@SiO₂ nanocomposite and cross-linked on the ITO magnetism-electrode with nafion (Nf) and glutaraldehyde (GA). The modified electrode was designated as Nf-GOD-HRP/Au-Fe₃O₄@SiO₂/ITO. The effects of some experimental variables such as the pH of supporting electrolyte, enzyme loading, the concentration of the mediator methylene blue (MB) and the applied potential were investigated. The electrochemical behavior of the biosensor was studied using electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and chronoamperometry. Under the optimized conditions, the biosensor showed a wide dynamic range for the detection of glucose with linear ranges of 0.05-1.0 mM and 1.0-8.0 mM, and the detection limit was estimated as 0.01 mM at a signal-to-noise ratio of 3. The biosensor exhibited a rapid response, good stability and anti-interference ability. Furthermore, the biosensor was successfully applied to detect glucose in human serum samples, showing acceptable accuracy with the clinical method.     

Quartz crystal microbalance and electrochemical cytosensing on a chitosan/multiwalled carbon nanotubes/Au electrode

The piezoelectric quartz crystal microbalance (QCM) was used to monitor the adhesion of mammalian cells on a chitosan (CS)/multiwalled carbon nanotubes (MWCNTs) composite modified gold electrode. The morphology and chemical properties of the CS/MWCNTs film were characterized with scanning electron microscopy (SEM) and Fourier transform infrared (FT-IR) spectroscopy. The human breast cancer cells (MCF-7) were adhered to and grown on the CS/MWCNTs film modified gold surface or a net CS film modified gold surface, and the process of which was continuously monitored and displayed by changes of the resonant frequency (Δf₀) and the motional resistance (ΔR₁) of the QCM. The attachment/spreading process of the MCF-7 cells on the QCM Au electrode decreased the Δf₀ and increased the ΔR₁ simultaneously, implying rather complicated effects (simultaneous mass, viscoelasticity and probable surface-stress load) on the sensor surface. The attachment rate and viability of the cells when proliferating on the two surfaces were detected by the MTT assay. The presence and state of cells on the electrode surface were confirmed by the fluorescent microscopy. Cyclic voltammetry and electrochemical impedance spectroscopy of the ferricyanide/ferrocyanide couple were examined before and after the cell adhesion. All data showed that the cell adhesion and proliferation processes were more efficient on the biocompatiable nanocomposite surfaces. The cell-based biosensor has potential for identification and screening of biologically active drugs and other biomolecules affecting cellular shape and attachment.     

Parylene-based implantable Pt-black coated flexible 3-D hemispherical microelectrode arrays for improved neural interfaces

In implantable medical systems, low-impedance electrode-tissue interface is important for maintaining signal quality for recording and effective charge transfer for stimulation. In this paper, we propose a novel hemispherical biocompatible and flexible microelectrode arrays (MEAs) which were fabricated by the process of micro electrical mechanical system (MEMS). Compared with conventional planar microelectrodes, the interface impedance of hemispherical microelectrodes decreased due to their increased surface area. Parylene C thin film with good biocompatibility and flexibility was chemical vapor deposited as packaging material for decreasing nerve tissue damage. Pt-black coatings were electroplated by applying current pulses in H₂PtCl₆ solution on electrode sites for the further decrease of interface impedance. Moreover, the geometrical and electrical properties of these MEAs were demonstrated by using a scanning electron microscope (SEM) and an electrochemical workstation. Experimental results showed that the interface impedance decreased by about 34% compared with conventional planar microelectrodes, and significantly decreased by 84% with Pt-black coatings on electrode sites compared with those uncoated microelectrodes.     

碳纳米复合材料在电化学生物传感器中的研究进展

碳纳米材料以其优异的导电特性和机械性能及极佳的生物相容性在构建电化学生物传感器中备受关注,为电化学生物传感器的开发和研究开辟了一片广阔天地。将碳纳米材料与其它纳米材料复合,是一种拓展和增强其应用的有效方法。碳纳米材料在电化学生物传感器方面的应用主要是作为传感器界面的修饰材料、生物分子的固载基质以及信号标记物等。该文综述了碳纳米复合材料在电化学生物传感器中的应用,包括碳纳米管纳米复合物、石墨烯纳米复合物、富勒烯及碳量子点纳米复合物。并展望了未来基于碳纳米材料的电化学生物传感器的研究方向。     

基于树状高分子的DNA电化学传感器对禽流感病毒的检测

将G4 PAMAM固定在玻碳电极表面,然后通过共价作用固定禽流感病毒探针ssDNA-1,以[Co(phen)3]3+为指示剂,采用示差脉冲伏安法和交流阻抗法对DNA电化学生物传感器进行了表征.结果发现,通过与双链dsDNA作用的[Co(phen)3]3+的峰电流信号的变化,可以识别和定量检测溶液中互补的禽流感病毒DNA片段.经过条件优化,该法测定DNA的浓度线性范围为1.3×10-9～6.5×10-8 mol/L,最低检测限为9.2×10-10 mol/L.