毛细管电泳安培法分析单个神经细胞  

Single Nerve Cell Analysis by Capillary Electrophoresis with Amperometric Detection

作　　者：胡深[1] 庞代文[1] 王宗礼[1] 程介克[1] 李之望[1] 樊友珍[1] 胡宏镇[1] 

机构地区：[1]武汉大学化学系,同济医科大学实验医学研究中心

出　　处：《高等学校化学学报》1996年第8期1207-1209,共3页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金

摘　　要：毛细管电泳安培法分析单个神经细胞胡深，庞代文，王宗礼，程介克（武汉大学化学系，武汉，４３００７２）李之望，樊友珍，胡宏镇（同济医科大学实验医学研究中心，武汉，４３００３０）关键词毛细管电泳，安培法检测，单个细胞分析在生命科学研究中，单个水平的神经细胞...This Paper reports the use of capillary electrophoresis (CE)with electrochemical detection to first detect the amol amount of norepinephrine (NE) and epinephrine(E) in a single sympathetic nerve cell (diameter 15￣25μm ,volume 2￣5pL) of rat.The home-made CE with electrochemical detection system consists of whole cell sampling, CE and amperometric detection. The whole cell sampling involves a micromanipulator, an inverted microscope and an in jection end of the capillary. To aid cell injection, the injection end of the capillary was etched to 100μm with hydrofluoric acid. The CE apparatus consists of 30 kV dc power supply , platinum wires and a fused-silica capillary (25μm i.d., 375μm i.d.,75 cm length) filled with 25 mmol/L MES buffer solution at pH 6.5.The amperometric detector used in this experiment was a carbon fiber microelectrode, The carbon fiber having a diame ter of 8μm and a length of 0.3mm,was inserted directly into the end of the electrophoresis capillary by micropositioner and microscope,The potential on the carbon fiber working electrode was controlled to 0.7V vs. SCE reference electrode by a potentiostat.Currents produced were measured and recorded by a picoammeter and a recorder.Sympathetic ganglia were dissected from rats. After enzymolysis the cells were isolated. The injection end of the capillary was placed adjacent to the cell using micromanipulator under the microscope,The whole cell sampling was accomplished for CE by directly drawing a cell into the tip of a capillary with electroosmotic flow and lysing the cell inside the capillary as observed under the microscope, using a micromanipulator, All separations were performed in 25 mmol MES buffer solution using 18 kV separation potential.The amperometric detector with microelectrode was used for detecting the neurotransmitters. The results in separation and detection the amol amount of NE and E in a single sympathetic nerve cell using this system are satisfactory.

关 键 词：毛细管电泳 安培法 神经细胞 

分 类 号：Q245[生物学—细胞生物学] O658.9[理学—分析化学]