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通过毛细管电泳结合电穿孔进行细胞内衍生化及激光诱导荧光检测对单个人类红细胞中的氨基酸进行分析。

Analysis of amino acids in individual human erythrocytes by capillary electrophoresis with electroporation for intracellular derivatization and laser-induced fluorescence detection.

作者信息

Zhang Hua, Jin Wenrui

机构信息

School of Chemistry and Chemical Engineering, State Key Laboratory of Microbial Technology, Shandong University, Jinan, China.

出版信息

Electrophoresis. 2004 Feb;25(3):480-6. doi: 10.1002/elps.200305642.

Abstract

A method for monitoring amino acids in single erythrocytes is described. For intracellular derivatization, reagent fluorescein isothiocyanate (FITC) was introduced into living cells by electroporation. For an 8 microm erythrocyte, the analytes were diluted by a factor of only 1.6. After completion of the derivatization reaction, a single cell was injected into the separation capillary tip and lysed there. The derivatized amino acids were separated by capillary electrophoresis, followed by laser-induced fluorescence detection. Nine amino acids were quantitatively determined, with amounts of amino acids ranging from 3.8-32 amol/single cell.

摘要

描述了一种监测单个红细胞中氨基酸的方法。对于细胞内衍生化,通过电穿孔将试剂异硫氰酸荧光素(FITC)引入活细胞。对于直径8微米的红细胞,分析物仅被稀释了1.6倍。衍生化反应完成后,将单个细胞注入分离毛细管尖端并在那里裂解。衍生化的氨基酸通过毛细管电泳分离,随后进行激光诱导荧光检测。定量测定了9种氨基酸,单个细胞中氨基酸的含量范围为3.8 - 32 amol。

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