CN113588950B - 一种单细胞内抗原的无线电化学可视化分析方法 - Google Patents

一种单细胞内抗原的无线电化学可视化分析方法 Download PDF

Info

Publication number
CN113588950B
CN113588950B CN202110686537.8A CN202110686537A CN113588950B CN 113588950 B CN113588950 B CN 113588950B CN 202110686537 A CN202110686537 A CN 202110686537A CN 113588950 B CN113588950 B CN 113588950B
Authority
CN
China
Prior art keywords
cell
cells
electrochemical
antigen
ecl
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202110686537.8A
Other languages
English (en)
Other versions
CN113588950A (zh
Inventor
江德臣
王玉玲
徐静娟
陈洪渊
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nanjing University
Original Assignee
Nanjing University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nanjing University filed Critical Nanjing University
Priority to CN202110686537.8A priority Critical patent/CN113588950B/zh
Publication of CN113588950A publication Critical patent/CN113588950A/zh
Application granted granted Critical
Publication of CN113588950B publication Critical patent/CN113588950B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G01N33/57415—Specifically defined cancers of breast
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
    • G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/49—Systems involving the determination of the current at a single specific value, or small range of values, of applied voltage for producing selective measurement of one or more particular ionic species
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57473—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving carcinoembryonic antigen, i.e. CEA
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • G01N33/57496—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving intracellular compounds
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/583—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with non-fluorescent dye label
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/902—Oxidoreductases (1.)
    • G01N2333/90245—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Urology & Nephrology (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • Physics & Mathematics (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Oncology (AREA)
  • Hospice & Palliative Care (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Electrochemistry (AREA)
  • Nanotechnology (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

电化学显微镜在过去的几十年中逐渐发展起来，用于单细胞生物分子的成像；然而，在单个细胞内实现胞内蛋白的电化学可视化是很困难的。本申请首次使用无线双极电化学发光技术(BPE‑ECL)观察了MCF 7细胞细胞核内的一个模型蛋白(KDM1/LSD1抗原)。亚微米尺寸的单壁碳纳米管与抗体相连接，用于识别相应的KDM1/LSD1抗原。在1000V/cm的低电场下，L012(鲁米诺类似物)在纳米管的阳极端被电化学氧化，发射出ECL信号，以实现其位置的无线可视化。在使用足够低的外加电压的情况下，可以观察到单个细胞内细胞核上的双极ECL发射，支持单个细胞内KDM1/LSD1抗原的电化学成像。

Description

一种单细胞内抗原的无线电化学可视化分析方法
技术领域
本申请属于电化学发光领域，具体涉及一种单细胞内抗原的无线电化学可视化分析方法。
背景技术
单细胞内蛋白质的特异性检测对于阐明细胞生物学的分子机制和疾病诊断具有重要意义。
荧光成像是一种常用的检测质膜和活细胞内部蛋白质的可视化方法。然而，背景荧光的存在影响了这些蛋白质的定量分析。电化学发光(electrochemiluminescence,ECL)作为一种近零背景的光学技术，被广泛应用于血清和细胞中各种抗原灵敏和定量的检测。在此过程中，发光基团(如Ru(bpy)3 2+)被标记在与相应抗原特异性结合形成免疫复合物的抗体上。在一定电压下，电极表面附近的发光体被激发产生中间体，中间体进一步与共反应物反应发射出ECL信号。然后通过显微镜和科研相机记录ECL信号，实现了抗原在单细胞质膜上的可视化。尽管ECL成像技术有了巨大的发展，但仍需要细胞与电极直接接触，这导致在单细胞内观察胞内蛋白很困难。因此，开发一种无线的电化学方法来实现单个细胞内蛋白质的可视化是电化学分析领域的一项长期任务。
双极电化学(BPE)是一种成熟的无线方法，它可在足够高的电场中使没有导线连接的导电物体产生极化，从而诱导了物体两端发生相应的电化学反应。这种方法不需要电极与细胞直接接触，允许细胞内的无线电分析。近年来，BPE-ECL已成功地用于分析生物分子和细胞表面的蛋白质。然而，BPE-ECL在单细胞内的进一步应用仍然具有挑战性。
原则上，要在一个细胞内实现BPE分析，导电物体必须小到亚微米尺度，这样它才能进入到细胞中。然而BPE有一个物理限制：即导体越短，所需施加的电压越大，才能使其极化并诱导ECL反应。理论计算得到，要在亚微米级的导体上诱导BPE-ECL反应，必须施加数万伏甚至更高的电压，这个电压不易操作。此外，这种高电压还会导致剧烈的水电解，在细胞介质中形成大量气泡，影响成像过程。
赖氨酸特异性去甲基化酶1(也可称为KDM1/LSD1)高表达会阻碍细胞分化，导致急性髓系白血病预后不良。
发明内容
为解决现有技术的上述问题，本申请提供一种单细胞内抗原的无线可视化分析方法，利用BPE-ECL实现了单个细胞内蛋白质的无线电化学可视化。
本申请提供一种单细胞内抗原的无线可视化分析方法，包括以下步骤：
首先，将单臂碳纳米管标记在抗体上，将标记后的抗体与细胞共同孵育，使标记后的抗体进入细胞内，并与胞内对应的抗原结合；
然后，用多聚甲醛将细胞固定，用曲拉通X-100使细胞通透；
最后，将处理好的细胞包埋在含有发光试剂和缓冲溶液的琼脂糖水凝胶内
此时，水凝胶中的混合液可通过通透的细胞膜进入到细胞内，通过馈电级在凝胶内施加电压，诱导细胞内的发光试剂在单壁碳纳米管上发生双极电化学发光反应，最终成像，实现细胞内抗原可视化。
在一实施例中，所述抗体为KDM1/LSD1抗体或癌胚抗原的抗体。
在一实施例中，所述细胞为人乳腺癌细胞。
在一实施例中，发光试剂为鲁米诺或其类似物。
在一实施例中，所述电压为1kV/cm～1.2kV/cm。
在一实施例中，所述馈电级为石墨电极
在一实施例中，所述电化学双极电化学发光反应成像的曝光时间为30s。
本申请还提供了一种单细胞内抗原的无线可视化分析方法，在检测单细胞内抗原中的应用。
本申请还提供了一种根据单细胞内抗原的无线可视化分析方法制备的试剂盒。
有益效果
本申请提供的这种在很低的电压下，可使双极电极发生局部增强极化。这个电压已被证明，要比经典的双极电极低至超过2个数量级。因此，利用一个较低的电压就足以使细胞内单个纳米管发生双极ECL反应。
使用L012(一种鲁米诺类似物，在低电压下具有强ECL发射)作为发光体，观察到细胞内纳米管上明亮的ECL信号，这是第一次在单细胞水平上实现细胞内蛋白的无线电化学可视化分析。本申请的技术方案解决了一个长期的挑战，即单个细胞内蛋白的电化学可视化，这将推动单细胞电化学分析的发展。
细胞首先与KDM1/LSD1抗体修饰的SWCNTs复合物共孵育，然后用多聚甲醛固定。最后，用曲拉通X-100使细胞通透，方便L012和PBS进入。
一般情况下，要在亚微米导体上诱导BPE-ECL反应，必须施加数万伏甚至更高的电压。同时，这种高电压导致剧烈的水电解会在细胞介质中形成大量的气泡，影响成像过程。在本申请中，为了避免气泡的产生，将导体包埋在含有电解质溶液的琼脂糖水凝胶中，在微米大小的导体上诱导BPE-ECL。水凝胶的多孔结构抑制了馈电极上气泡的形成，提供了微小导体上实现无线的ECL可视化的解决方案。
由于SWCNTs具有良好的细胞通透性和生物相容性，SWCNTs-抗体复合物可被载入细胞并与细胞内抗原结合，如图1所示。然后将固定的细胞包埋到水凝胶中，以避免在BPE-ECL过程中出现气泡。更重要的是，由于凝胶内空间上限制，电压降可以发生在保留细胞的水凝胶的微孔内。
附图说明
图1.用于细胞核上KDM1/LSD1抗原的无线电化学可视化分析的装置示意图。放大的部分显示了L012在单个SWCNTs上的双极ECL反应，该SWCNTs是通过抗体-抗原免疫复合物被标记在细胞核上。
图2.(A)L012在SWCNTs-COOH修饰电极上的循环伏曲线(红色线)和ECL曲线(黑色线)。溶液是含有1mM L012的10mM PBS(pH 7.4)。扫描速率为10mV/s。(B)毛细管内水凝胶中的SWCNT的ECL图像。曝光时间为30秒。
图3.用CEA抗体/SWCNTs标记的MCF 7细胞的(A)明场图像、(B)ECL图像和(C)两者的叠加图像。ECL成像的曝光时间为30s。
图4.KDM1/LSD1抗体修饰的SWCNT被加载到细胞内并与细胞核结合的(A)明场图像、(B)ECL图像和(C)两者的叠加图像。ECL成像的曝光时间为30s。
图5：Alexa488与SWCNTs标记的抗体的偶联复合物孵育MCF 7细胞的明场(A)荧光图像(B)和两者的叠加图像(C)。曝光时间为1s。
具体实施方式
以下内容仅例示发明的原理。因此，虽未在本说明书中明确地进行说明或图示，但本领域技术人员可实现发明的原理而发明包括在发明的概念与范围内的各种装置。另外，应理解，本说明书中所列举的所有附有条件的术语及实施例在原则上仅明确地用于理解发明的概念，并不限制于像这样特别列举的实施例及状态。
实施例一水凝胶中单臂碳纳米管的可视化实验
为了使用BPE-ECL对单个SWCNTs进行初始可视化，将含有SWCNTs-COOH(0.005％wt)、L012(鲁米诺类似物)(1mM)和10mM磷酸盐缓冲液(PBS,pH 7.4)的琼脂糖水凝胶放置在毛细管(id.0.6mm)中。通过透射电子显微镜，发现单个SWCNTs-COOH的长度小于500nm，宽度则小于10nm。在毛细管内形成凝胶后，SEM图像展示出：凝胶为孔径小于70μm的多孔结构，适合保留一个细胞。
根据本实施例的结果可知，当只施加了1kV/cm的电场时，在毛细管中就能观察到可分辨的ECL管板，参见图2B。随着电场从1kV/cm逐渐增大到1.2kV/cm，这些点的ECL强度逐渐增强。ECL强度与外加电场的依赖关系表明，ECL反应确实发生在SWCNTs上。此外，每个ECL点的尺寸在0.5～1μm之间，与一个纳米管的尺寸相匹配。表征出SWCNTs上的电化学反应所需的最小电压，为1kV/cm。
实施例二单个细胞膜上癌胚抗原的可视化标记
纳米管用于单细胞BPE-ECL成像的在细胞膜上显示抗原。癌胚抗原的抗体(anti-CEA Ab)被标记在SWCNTs上后形成anti-CEA Ab-SWCNTs复合物，再与固定的MCF 7细胞(该细胞的细胞膜上高表达CEA抗原)一起孵育，然后将其包埋在含有L012的琼脂糖水凝胶里，在两个馈电极上施加1000V电压，同时记录ECL成像。从图3A-C可以观察到，单个细胞的整个表面都产生了可见的ECL光斑，且与明场的细胞有很好的重叠。此外，为了排除可能存在的SWCNTs的非特异性吸附，将细胞与不含抗体的SWCNTs孵育，并收集ECL图像。结果显示细胞上几乎没有观察到ECL光斑，证明SWCNTs在细胞中的非特异性吸附可忽略不计。以上结果充分的证明BPE-ECL方法可以用于单个细胞的膜蛋白的无线可视化分析。
实施例三细胞核上KDM1/LSD1蛋白的可视化标记
本申请的进一步进行了可视化分析细胞内的抗原的实验，即：赖氨酸特异性去甲基化酶1(也称为KDM1/LSD1)。KDM1/LSD1高表达会阻碍细胞分化，导致急性髓系白血病预后不良。首先，细胞先与KDM1/LSD1抗体修饰的SWCNTs共孵育，然后用多聚甲醛固定。最后用曲拉通X-100使细胞通透，方便L012和缓冲液进入。然后，在馈电极上施加1kV/cm的电压，记录ECL图像。如图4A和B中所示，明场和ECL图像的重叠图像(图4C)表明ECL发射都集中在细胞内的细胞核附近，且与膜蛋白的ECL可视化分布明显不同(图3C)。再结合抗体修饰的SWCNTs位置的荧光表征(图5)，充分反映了细胞内细胞核中KDM1/LSD1抗原的信息。
为了排除SWCNTs的非特异性吸附带来的假阳性信号，选择用不含修饰抗体的纯SWCNTs与细胞共培养，进行了对照实验。发现微弱的ECL斑点几乎遍布整个细胞，且不针对任何细胞室。实验结果说明SWCNTs虽然进入了细胞内，但由于没有抗体，无法被特异性结合在细胞核上，进一步证实抗体修饰的纳米管可以有效地与目标蛋白所在特定的胞腔内结合，实现无线电化学可视化。
如上所述，参照本发明的优选实施例进行了说明，但本技术领域内的普通技术人员可在不脱离随附的权利要求书中所记载的本发明的思想及领域的范围内对本发明进行各种修正或变形而实施。

Claims (6)

1.一种单细胞内抗原的无线可视化分析方法，其特征在于，包括以下步骤：
首先，将单臂碳纳米管标记在抗体上，将标记后的抗体与细胞共同孵育，使标记后的抗体进入细胞内，并与胞内对应的抗原结合；
然后，用多聚甲醛将细胞固定，用曲拉通X-100使细胞通透；
最后，将处理好的细胞包埋在含有发光试剂和缓冲溶液的琼脂糖水凝胶内，水凝胶中的混合液可通过通透的细胞膜进入到细胞内，通过馈电级在凝胶内施加电压，诱导细胞内的发光试剂在单壁碳纳米管上发生双极电化学发光反应，最终成像，实现细胞内抗原可视化；
其中，所述发光试剂为L012；所述电压为1 kV/cm~1.2 kV/cm；其中，单个SWCNTs-COOH的长度小于500 nm，宽度则小于10 nm；凝胶为孔径小于 70 μm的多孔结构，适合保留一个细胞。
2.根据权利要求1所述的方法，其特征在于，所述抗体为KDM1/LSD1抗体或癌胚抗原的抗体，所述方法不以获得疾病和/或健康状况为直接目的。
3.根据权利要求1所述的方法，其特征在于，所述细胞为人乳腺癌细胞，所述方法不以获得疾病和/或健康状况为直接目的。
4.根据权利要求1所述的方法，其特征在于，所述馈电级为石墨电极。
5.根据权利要求1所述的方法，其特征在于，所述电化学双极电化学发光反应成像的曝光时间为30s。
6.一种根据权利要求1所述的单细胞内抗原的无线可视化分析方法，在检测单细胞内抗原中的应用。
CN202110686537.8A 2021-06-21 2021-06-21 一种单细胞内抗原的无线电化学可视化分析方法 Active CN113588950B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110686537.8A CN113588950B (zh) 2021-06-21 2021-06-21 一种单细胞内抗原的无线电化学可视化分析方法

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110686537.8A CN113588950B (zh) 2021-06-21 2021-06-21 一种单细胞内抗原的无线电化学可视化分析方法

Publications (2)

Publication Number Publication Date
CN113588950A CN113588950A (zh) 2021-11-02
CN113588950B true CN113588950B (zh) 2023-07-18

Family

ID=78244149

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110686537.8A Active CN113588950B (zh) 2021-06-21 2021-06-21 一种单细胞内抗原的无线电化学可视化分析方法

Country Status (1)

Country Link
CN (1) CN113588950B (zh)

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4336812B2 (ja) * 2003-03-07 2009-09-30 博夫 岩田 細胞に発現する表面抗原を迅速に同定するための分析方法
FR3019897A1 (fr) * 2014-04-11 2015-10-16 Univ Bordeaux Procede de generation de lumiere par electro-chimiluminescence par une dispersion de particules electriquement conductrices dans un solvant
CN105928927B (zh) * 2016-04-15 2019-03-12 南京大学 电致化学发光成像方法快速分析单细胞内成分的方法

Also Published As

Publication number Publication date
CN113588950A (zh) 2021-11-02

Similar Documents

Publication Publication Date Title
Wu et al. Visual electrochemiluminescence detection of cancer biomarkers on a closed bipolar electrode array chip
Sentic et al. 3D electrogenerated chemiluminescence: from surface-confined reactions to bulk emission
Wang et al. Facile synthesis of cuprous oxide nanowires decorated graphene oxide nanosheets nanocomposites and its application in label-free electrochemical immunosensor
Barker et al. Scanning electrochemical microscopy: beyond the solid/liquid interface
Jie et al. Quantum dots-based multifunctional dendritic superstructure for amplified electrochemiluminescence detection of ATP
Zhang et al. Signal-on electrochemiluminescent immunosensor based on poly (amidoamine) dendrimer functionalized carbon nanodots amplification for ultrasensitive detection of α-fetoprotein
Liao et al. Reagentless electrochemiluminescent detection of protein biomarker using graphene-based magnetic nanoprobes and poly-L-lysine as co-reactant
Liu et al. Application of ZnO quantum dots dotted carbon nanotube for sensitive electrochemiluminescence immunoassay based on simply electrochemical reduced Pt/Au alloy and a disposable device
Liao et al. Amplified electrochemiluminescent immunosensing using apoferritin-templated poly (ethylenimine) nanoparticles as co-reactant
Ding et al. Switches-controlled bipolar electrode electrochemiluminescence arrays for high-throughput detection of cancer biomarkers
Zhang et al. An ITO bipolar array for electrochemiluminescence imaging of H2O2
Yin et al. A highly sensitive electrochemical IFN-γ aptasensor based on a hierarchical graphene/AuNPs electrode interface with a dual enzyme-assisted amplification strategy
Xie et al. A novel binary luminophore based high-efficient electrochemiluminescence biosensor for ultrasensitive detection of human epidermal growth factor receptor-2
US10655121B2 (en) Galvanotaxis assay for quantitative assessment of metastatic potential of cancer cells
US20220364991A1 (en) Method and System for Decoding Information Stored on a Polymer Sequence
Liu et al. Electrochemiluminescent biosensor with DNA link for selective detection of human IgG based on steric hindrance
CN113588950B (zh) 一种单细胞内抗原的无线电化学可视化分析方法
Zhang et al. Enhanced electrochemiluminescence at single lithium iron phosphate nanoparticles for the local sensing of hydrogen peroxide efflux from single living cell under a low voltage
Cao et al. Enzyme-loaded liposome with biocatalytic precipitation for potentiometric immunoassay of thyroid-stimulating hormone in thyroid carcinoma
Wang et al. Wireless electrochemical visualization of intracellular antigens in single cells
Li et al. Ultrasensitive multiplexed protein biomarker detection based on electrochemical tag incorporated polystyrene spheres as label
Liu et al. Nanovehicles based bioassay labels
Gao et al. Enhanced electrochemiluminescence cytosensing based on abundant oxygen vacancies contained 2D nanosheets emitter coupled with DNA device cycle-amplification
Li et al. A nonenzymatic electrochemical immunosensor for ultrasensitive detection of tumor biomarkers based on palladium nanoparticles conjugated reduced graphene nanosheets
CN108088882A (zh) 干细胞的电化学检测方法

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant