超微电极实时监测植物细胞壁参与调控的单细胞活性氧爆发（英文）

植物细胞活性氧爆发在植物的抗病以及信号转导中起着非常重要的作用,植物内活性氧产生及代谢受到复杂而精确的机制调控,从而维持正常的活性氧水平以发挥其生理功能.然而,在单细胞水平开展活性氧爆发实时监测及其调控机制研究一直受到很大的挑战.本文以碳纤维微盘电极(CFMDE)为基底电极,利用Nafion的模板效应,采用电化学沉积法制得纳米铂颗粒修饰电极(NPt/Nafion/CFMDE);同时采用基于聚二甲基硅氧烷(PDMS)的软光刻技术,制备了一种高效固定植物悬浮细胞的琼脂糖阵列微孔芯片.使用NPt/Nafion/CFMDE实时监测了单个拟南芥原生质体活性氧爆发,并证明电化学监测活性氧的主要成分为过氧化氢.在此基础上,采用浅层培养法培养原生质体再生植物细胞壁.电化学监测结果表明,与单个原生质体相比,植物细胞在受到刺激时释放的过氧化氢量显著降低;然而当采用过氧化物酶抑制剂抑制植物细胞壁上过氧化物酶活性后,植物细胞释放过氧化氢量显著回升.研究结果表明,细胞壁在活性氧爆发过程具有很好的调控功能,可望促进植物细胞活性氧爆发及其调控机制的研究.     

微流控芯片操纵传输及实时监测单细胞量子释放

微流控芯片技术用于细胞生化分析已引起了广泛关注.Harrison等首次在微流控芯片上对细胞群体进行操纵、传输及反应.yang等在微流控芯片上操纵细胞群体的排列,并用荧光检测细胞群体摄取钙的反应.至今还未见到微流控芯片对单个细胞进行操纵传输、定位及实时监测的报道.单细胞受激释放的监测对探索生物体神经传导具有重要意义.     

超微电极电化学及显微荧光成像时空监测单细胞释放

细胞的受激释放即分泌化学信号分子是胞间通讯最重要、最广泛的一种途径,是其参与生命活动的重要形式,在生命活动中起着非常重要的作用.对其进行研究在神经生物学、细胞生物学、病理学和药学等多个学科领域中都具有非常重要的意义.     

微纳电化学传感界面的构建及其用于单细胞实时监测

细胞是生物体形态结构和生命活动的基本单位.常规检测群体细胞的方法往往会掩盖细胞间的个体差异,因此亟需发展高效的单细胞分析策略,深入研究细胞生命活动过程,揭示疾病发生发展机制,推动个体化诊疗.超微电化学传感器具有尺寸小、灵敏度高、时空分辨率高等特点,在单细胞实时动态监测方面发挥了非常重要的作用.目前,微纳电化学传感器在电...     

光致清洁电极可见光在线更新与细胞实时监测

电化学传感器在用于细胞实时监测过程中,电极界面污染严重影响其检测性能.通过将纳米光催化剂与电化学传感材料复合,构建光致清洁电化学传感器,为电极界面的高效及无损更新提供了新思路.然而光催化产生的活性氧自由基导致细胞损伤,限制了细胞培养及检测过程中电极界面的实时更新.为此,我们在PEDOT@CdS/TiO₂/ITO可见光致更新电极表面旋涂明胶薄层,在保持电极良好的光致清洁和电化学传感性能同时,利用明胶薄层阻碍光催化产生的活性氧自由基扩散至细胞表面,显著降低了细胞损伤.此外,明胶优良的生物相容性有利于细胞的黏附及增殖.利用该电极,我们实现了人脐静脉内皮细胞（HUVECs）培养过程中,电极的在线更新以及细胞释放一氧化氮的实时监测.     

光纤动态过程实时监测系统的研制

一、引言光导纤维技术及其应用是当今发展十分迅速的一个研究领域。光导纤维作为一种传输光信号的工具,具有损耗低、抗干扰强、信息承载量大等优点。另外,光纤探头很容易实现微型化,加之其良好的柔软性能和机械性能,为实现原位检测、遥测以及过程控制提供了一种强有力的手段。近年来,大量基于不同机理的光纤传感器的涌现,展示了该技术在分析、检测     

质谱实时监测有机化学反应的研究进展

质谱法是一种能够提供物质丰富结构信息的谱学方法,广泛应用于有机物的测定、有机化学反应的监测以及有机化学反应机理的研究中。本文综述了质谱实时监测缩合反应、取代反应、氧化反应、环化反应、加成反应、偶联反应、自由基反应等有机化学反应的研究进展。     

肾小球滤过率活体实时监测荧光示踪剂的研究进展

急性肾损伤是一种临床常见的急性疾病. 通过外源性的荧光示踪剂对肾小球滤过率进行非侵入性的实时监测, 对于及时了解易患病人的肾脏功能具有重要的临床价值和应用前景. 本文综合评述了用于活体检测肾小球滤过率的外源性荧光示踪剂的近期研究进展, 重点介绍了荧光示踪剂的设计策略及其活体成像应用效果, 并对相应检测装置的发展进行了阐述, 同时对该领域的挑战与发展前景进行了展望.     

分子信标用于核酸连续复制过程的体外实时监测

利用分子信标核酸探针实时监测了核酸连续复制过程. 分子信标不仅作为模板参与复制反应, 而且同步将复制过程的信息转换为荧光信号, 实现复制过程的体外实时监测. 该方法不仅为DNA复制检测提供了一种实时研究手段, 而且为核酸复制动力学及与复制相关疾病的深入研究提供了一种新的思路.     

基于分子信标实时监测大肠杆菌DNA连接酶催化的DNA连接过程

报道了一种对DNA连接过程进行实时监测的方法,利用分子信标核酸探针作为DNA连接反应的模板和检测探针,实时监测了 E.coli DNA连接酶催化的DNA连接反应,克服了传统的凝胶电泳技术操作复杂、周期长及无法实时监测DNA连接过程的缺点,为核酸连接过程的实时监测和连接酶催化机理的研究提供了更为丰富的信息.在此基础上,发展了一种快速、准确测定 E.coli DNA连接酶的方法,线性响应范围为4.0×10^-6～2.0×10^-4U/μL,检测下限为4.0×10^-6U/μL.     

单细胞水平的分析方法研究及进展

本文评述了近年来单细胞分析的应用及进展，介绍了超微电极电化学分析法和微柱分离法两大类方法在单细胞分析中应用的机理、必要的技术、有关研究内容和最新进展及其发展方向。     

微流控芯片电泳/电化学检测人单个肝癌细胞中的谷胱甘肽

采用微流控装置结合电化学检测研究了测定人单个血红细胞中谷胱甘肽(GSH)的方法。在该方法中,细胞的进样、定位、溶膜以及细胞中谷胱甘肽的转移和检测都在配有通道端安培检测器的双T形芯片中完成。单个细胞用液压导入到双T的交界面,在电泳缓冲液中毛地黄皂苷的作用下,细胞膜被穿孔。再施加直流电压,细胞被溶膜。释放出来的GSH被此直流电压电迁移至通道端并在Au/Hg电极上被检测。用校正曲线法可以定量测定单个细胞中的GSH。     

Analysis of Amino Acids in a Single Human Red Blood Cell by Capillary Zone Electrophoresis with Intracellular NDA—derivatization and Electrochemical Detection

A novel method for determination of amino acids in individual red blood cells has been developed. In this method, the derivatization reagents (NDA and CN^-) are introduced into living cells by electroporation. After completion of derivatization,the amino acids in a single cell is determined by capillary zone electrophoresis with end-column amperometric detection.     

Stretchable Electrochemical Sensor for Real‐Time Monitoring of Cells and Tissues

Stretchable electrochemical sensors are conceivably a powerful technique that provides important chemical information to unravel elastic and curvilinear living body. However, no breakthrough was made in stretchable electrochemical device for biological detection. Herein, we synthesized Au nanotubes (NTs) with large aspect ratio to construct an effective stretchable electrochemical sensor. Interlacing network of Au NTs endows the sensor with desirable stability against mechanical deformation, and Au nanostructure provides excellent electrochemical performance and biocompatibility. This allows for the first time, real‐time electrochemical monitoring of mechanically sensitive cells on the sensor both in their stretching‐free and stretching states as well as sensing of the inner lining of blood vessels. The results demonstrate the great potential of this sensor in electrochemical detection of living body, opening a new window for stretchable electrochemical sensor in biological exploration.     

Assay of Histamine in Single Mast Cells by Capillary Zone Electrophoresis with Electrochemical Detection

Histamine is one of the chemical mediators in connection with allergies. Only Pihel et al.1 reported a method for detection of histamine in isolated mast cells by high- performance liquid chromatography with electrochemical detection. In this method, single cells were removed and transferred to 300-nL microvials. A part of the supernatant from individual vials was injected into the chromatography column. The analysis of histamine in individual rat peritoneal mast cells using CZE with amp…     

毛细管电泳安培测定鼠单个交感神经细胞中儿茶酚胺类神经递质

用自组装的毛细管电泳微电极安培检测系统，分析了鼠的单个交感神经细胞。大白鼠经麻醉、微解剖及酶解分离出新鲜的单个交感神经细胞。采用全细胞进样，用异丙肾上腺素 作内标。测定了儿茶酚胺类神经递质去甲肾上腺素和肾上腺素。检测限分别达 ５７ ａｍｏｌ和 ５９ ａｍｏｌ。八个活细胞分析结果，每个细胞平均含去甲肾上腺素 ４５５ ａｍｏｌ及肾上腺素 ６５ ａｍｏｌ。     

Real‐Time Monitoring of Auxin Vesicular Exocytotic Efflux from Single Plant Protoplasts by Amperometry at Microelectrodes Decorated with Nanowires

Recent biochemical results suggest that auxin (IAA) efflux is mediated by a vesicular cycling mechanism, but no direct detection of vesicular IAA release from single plant cells in real‐time has been possible up to now. A TiC@C/Pt‐QANFA micro‐electrochemical sensor has been developed with high sensitivity in detection of IAA, and it allows real‐time monitoring and quantification of the quantal release of auxin from single plant protoplast by exocytosis.     

单细胞的毛细管电泳分析

对单细胞分析化学中毛细管电泳分离技术的进样方式、检测器、应用和前景予以综述,４６篇。     

Intracellular Wireless Analysis of Single Cells by Bipolar Electrochemiluminescence Confined in a Nanopipette

The inside walls of a nanopipette tip are decorated by a Pt deposit that is used as an open bipolar electrochemiluminescence (ECL) device to achieve intracellular wireless electroanalysis. The synergetic actions of nanopipette and of bipolar ECL lead to the spatial confinement of the voltage drop at the level of the Pt deposit, which generates ECL emission from luminol. The porous structure of Pt deposit permits the electrochemical transport of intracellular molecules into the nanopipette that is coupled with enzymatic reactions. Thus, the intracellular concentrations of hydrogen peroxide or glucose are measured in vivo as well as the intracellular sphingomyelinase activity. In comparison with the classic bipolar ECL, the remarkably low potential applied in our approach is restricted inside the nanopipette and it minimizes the potential bias of the voltage on the cellular activity. Accordingly, this wireless ECL approach provides a new direction for analysis of single living cells.     

Electrochemical Gene‐Function Analysis for Single Cells with Addressable Microelectrode/Microwell Arrays

To each his own : An addressable electrochemical device consisting of orthogonally arranged rows and columns of electrodes has been constructed to monitor protein expression in genetically engineered cells at the single‐cell level. The response based on redox cycling reflected the different expression levels of the enzyme from individual HeLa cells transfected with a plasmid vector including secreted alkaline phosphatase.