Zhang Hua, Jin Wenrui
School of Chemistry and Chemical Engineering, State Key Laboratory of Microbial Technology, Shandong University, Jinan, China.
Electrophoresis. 2004 Feb;25(3):480-6. doi: 10.1002/elps.200305642.
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.
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。