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微芯片电泳-化学发光检测法测定小鼠单个纤维肉瘤细胞中牛磺酸和氨基酸的含量。

Quantification of taurine and amino acids in mice single fibrosarcoma cell by microchip electrophoresis coupled with chemiluminescence detection.

机构信息

Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education), College of Chemistry and Chemical Engineering, Guangxi Normal University, Guilin, PR China.

出版信息

Electrophoresis. 2010 May;31(10):1630-6. doi: 10.1002/elps.200900665.

Abstract

A method based on MCE coupled with chemiluminescence (CL) detection was developed for the determination of taurine (Tau) and amino acids including alanine (Ala), glycine (Gly), tryptophan (Trp), glutamic acid (Glu) and aspartic acid (Asp) present in mice single fibrosarcoma (S180) cells. Cell injection, loading, cytolysis, electrophoretic separation and CL detection were integrated onto a simple double-T microfluidic chip. The intracellular constituents were electrophoretically separated within 150 s. The CL detection was based on the enhancement effects of Tau and amino acids on the CL reaction of luminol with H(2)O(2) and Cu(2+). The average amounts of Tau, Trp, Gly, Ala, Glu and Asp in per S180 cell from a cell population were 4.73, 1.23, 2.65, 1.94, 1.61 and 1.99 fmol. Ten S180 cells were analyzed, and the contents of Tau, Trp, Gly, Ala, Glu and Asp in mice single S180 cells were found to be in the range of 1.78-8.84, 0.95-2.31, 1.08-6.87, 1.03-4.05, 0.84-2.61 and 0.82-3.68 fmol, respectively. This work demonstrates that MCE coupled with CL detection is a useful analytical tool that is simple, quick and highly sensitive for single-cell analysis.

摘要

建立了一种基于微流控芯片毛细管电泳(MCE)与化学发光(CL)检测联用的方法,用于测定小鼠纤维肉瘤(S180)单个细胞内的牛磺酸(Tau)和丙氨酸(Ala)、甘氨酸(Gly)、色氨酸(Trp)、谷氨酸(Glu)和天冬氨酸(Asp)等氨基酸。将细胞注射、加载、细胞裂解、电泳分离和 CL 检测集成在一个简单的双 T 型微流控芯片上。细胞内成分在 150s 内实现电泳分离。CL 检测基于 Tau 和氨基酸对鲁米诺-H2O2-Cu2+化学发光反应的增强作用。从细胞群体中每个 S180 细胞中 Tau、Trp、Gly、Ala、Glu 和 Asp 的平均含量分别为 4.73、1.23、2.65、1.94、1.61 和 1.99 fmol。分析了 10 个 S180 细胞,发现单个 S180 细胞中 Tau、Trp、Gly、Ala、Glu 和 Asp 的含量范围分别为 1.78-8.84、0.95-2.31、1.08-6.87、1.03-4.05、0.84-2.61 和 0.82-3.68 fmol。该工作表明,MCE 与 CL 检测联用是一种简单、快速、高灵敏度的单细胞分析工具。

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