Na+- and Ca2+-selective microelectrodes were made with Simon's neutral carrier ETH 227 and ETH 1001, respectively, and their properties were studied for intracellular application. The kNaK (selectivity coefficient for Na+ with respect to K+) values of the Na+-selective microelectrodes were in the range of 0.01-0.02, which is comparable to those of recessed-tip Na+-selective glass microelectrodes. The kNaMg values of the microelectrodes were approximately 0.005 so that the interference by intracellular Mg2+ levels could be negligible. The kNaCa values were approximately 2 and the Na+-selective microelectrodes were more selective to Ca2+ than Na+. This indicates that their intracellular application requires special care to handle Ca2+ interference under certain conditions. The kNaK, kNaMg, and kNaCa values did not depend significantly on the methods used for their determination or on the ion activity levels tested. The Nicolsky equation described well the microelectrode potentials in the mixed solutions of NaCl (1-100 mM) and KCl. Potential and resistance of the microelectrodes were stable for a long period and their response time was fast. The results indicate that the Na+-selective microlectrodes are suitable for measurements of intracellular Na ion activities. Ca2+-selective microelectrode potentials at Ca2+ concentrations lower than 10(-4) M changed significantly for the first 2-3 h and then became fairly stable. The rate of the potential change was dependent on the column length of the Ca2+-selective liquid filled. Potentials of the microelectrodes varied from 10-20 mV for Ca2+ between 10(-7) and 10(-6) M concentrations, which may be the cytosolic free-Ca2+ range. With the Ca2+ concentrations greater than 10(-6) M, the microelectrodes had potential changes of approximately 30 mV or greater for a tenfold change in Ca2+ concentration. The kCaK and kCaNa values were in the ranges of 10(-5)-10(-6) and 10(-4)-10(-5), respectively. The kCaMg values were approximately 10(-7). The results show that the Ca2+-selective microelectrodes can be used for measurements of cytosolic Ca ion activities.
Electrochemical properties of Na+-selective glass microelectrodes were studied and compared with those of K+-selective glass microelectrodes. The selectivity of Na+-selective glass microelectrodes depended on the ion concentration of test solutions. With aging, resistance of Na+-selective microelectrodes increased and their selectivity for Na over K decreased. Na+-selective microelectrodes potential measured in NaCl solution remained constant with aging, while the potential measured in KCl solution decreased and became more positive. The changes in resistance and potential of Na+-selective microelectrodes may be due to the effects of the less mobile cation, i.e., H+ or K+ on the Na ion exchange in the Na-sensing region. The results indicate that Na+-selective microelectrodes must be used as soon after filling as possible. The selectivity of Na+-selective microelectrodes increased with increase of the sensitive exposed-tip length, whereas their response time became slow due to a large recessed volume, indicating requirement of an optimum exposed-tip length for intracellular applications. The changes in the properties of Na+-selective glass microelectrodes with aging contrasted with those of K+-selective glass microelectrodes in which resistance decreased and K+-selectivity increased. The K+-selective microelectrodes required aging before use for a high selectivity and low resistance. The K+-selective microelectrodes with low resistance after sufficient aging can be used without insulation to measure K+ and Na+ activities in aqueous solutions. The different properties between Na+- and K+-selective microelectrodes are understandable, because hydration of N+-selective glass is much less extensive than that of K+-selective glass.
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