Analysis of amino acids in individual human erythrocytes by capillary electrophoresis with electroporation for intracellular derivatization and laser-induced fluorescence detection.
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.
Zhang
Hua
H
School of Chemistry and Chemical Engineering, State Key Laboratory of Microbial Technology, Shandong University, Jinan, China.
Jin
Wenrui
W
D000596:Amino Acids / Q000032:analysis* / Q000737:chemistry; D003584:Cytological Techniques; D019075:Electrophoresis, Capillary / Q000379:methods*; D018274:Electroporation; D004912:Erythrocytes / Q000737:chemistry*; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005453:Fluorescence; D006801:Humans; D007834:Lasers; D015203:Reproducibility of Results
D000596:Amino Acids; D016650:Fluorescein-5-isothiocyanate