Microchip electrophoresis with chemiluminescence detection for assaying ascorbic acid and amino acids in single cells.
Microchip electrophoresis with chemiluminescence detection for assaying ascorbic acid and amino acids in single cells.
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DOI：
10.1016/j.chroma.2009.08.008
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发表时间：
2009-09-25
期刊：
影响因子：
--
通讯作者：
Liu YM
中科院分区：
文献类型：
--
作者：
Zhao S;Huang Y;Liu YM
建立了一种基于微芯片电泳（MCE）化学发光（CL）检测的单细胞抗坏血酸（AA）及色氨酸（Trp）、甘氨酸（Gly）、丙氨酸（Ala）等氨基酸的检测方法。细胞注射、上样、裂解、电泳分离和CL检测都集成在一个简单的交叉微流控芯片上。通过在储层上施加一组电位，电泳方法将单个细胞加载在交叉路口。对接的细胞在直接电场作用下迅速裂解。细胞内内容物在130s内进行MCE分离。CL检测是基于AA和氨基酸对鲁米诺与K3[Fe(CN)6] CL反应的增强作用。制备大鼠肝细胞作为实验细胞模型进行分析。单个大鼠肝细胞内AA、Trp、Gly和Ala的平均含量分别为38.3 fmol、5.15 fmol、3.78 fmol和3.84 fmol （n=12）。
A method based on microchip electrophoresis (MCE) with chemiluminescence (CL) detection was developed for the determination of ascorbic acid (AA) and amino acids including tryptophan (Trp), glycine (Gly) and alanine (Ala) present in single cells. Cell injection, loading, lysing, electrophoretic separation, and CL detection were integrated onto a simple cross microfluidic chip. A single cell was loaded in the cross intersection by electrophoretic means through applying a set of potentials at the reservoirs. The docked cell was lysed rapidly under a direct electric field. The intracellular contents were MCE separated within 130 s. CL detection was based on the enhancing effects of AA and amino acids on the CL reaction of luminol with K3[Fe(CN)6]. Rat hepatocytes were prepared and analyzed as the test cellular model. The average intracellular contents of AA, Trp, Gly and Ala in single rat hepatocytes were found to be 38.3 fmol, 5.15 fmol, 3.78 fmol and 3.84 fmol (n=12), respectively.
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