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Microchip electrophoresis with chemiluminescence detection for assaying ascorbic acid and amino acids in single cells

机译:微芯片电泳具有化学发光检测用于在单细胞中测定抗坏血酸和氨基酸

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摘要

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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