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采用钙离子敏感中性载体ETH 1001研制成外径<0.7 um的钙离子敏感微电极(Ca~(2+)—ISM),Ca~(2+)浓度在10~(-3)～10~(-7)M之间,平均斜率为25.4 mV。检测下限为10~(-8)M。应用该Ca~(2+)-ISM及常用微电极,同时测定了蟾蜍缝匠肌细胞内的钙离子活度α_(Ca)~i和细胞膜电位(E_M),测定结果为:α_(Ca)~i=0.11um,E_M=-79.0mV。通过分别改变任氏液中的钾离子浓度[K~+]。和钙离子浓度[Ca~(2+)]。测定α_(Ca)~i了及E_M的动态变化。结果表明:[K~+],从3.5 mM升高至10 mM时,α_(Ca)~i由0.11μM变为0.15 uM,E_M由-79.0mV变为-67.7 mV[Ca~(2+)],从1.8mM降低为1μM时,α_(Ca)~i由0.11 μM穸为0.03μM,E_M由-79.0mV变为-73.8mV。实验表明Ca~(2+)-ISM是细胞内钙离子动态检测极为有效,方便和直观的方法之一。
The calcium ion sensitive microelectrode (Ca2 +) - ISM with outer diameter <0.7 um was prepared by using ETH 1001 with Ca2 + sensitivity. The concentration of Ca2 + was 10-3 ~ (-7) M, with an average slope of 25.4 mV. The lower limit of detection is 10 ~ (-8) M. The Ca ~ (2 +) - ISM and common microelectrodes were used to measure the activity of Ca ~ (i) and cell membrane potential (E_M) in Toad sartorius muscle cells. The results were as follows: ~ i = 0.11um, E_M = -79.0mV. By changing the concentration of potassium in Ren liquid separately [K ~ +]. And calcium ion concentration [Ca ~ (2+)]. The dynamic changes of α_ (Ca) ~ i and E_M were measured. The results showed that when [K ~ +] increased from 3.5 mM to 10 mM, α_ (Ca) ~ i changed from 0.11 μM to 0.15 μM and E_M changed from -79.0 to -67.7 mV [Ca 2+] ], From 1.8 mM to 1 μM, α_ (Ca) -i was changed from 0.11 μM 0.03 to 0.03 μM, and E_M changed from -79.0 mV to -73.8 mV. Experiments show that Ca ~ (2 +) - ISM is one of the most effective, convenient and intuitive methods for detecting intracellular calcium dynamics.