Submicron tip breakage and silanization control improve ion-selective microelectrodes.

Probable causes of failure of otherwise well-constructed liquid ion-exchanger (LIE) micro-electrodes of average tip size less than 0.15 micron were examined. The problem could be attributed to two major variables, both localized at the tip: partial tip occlusion during fabrication prevents the generation of an electromotive force (small or absent slope and/or selectivity, high resistance); or poor hydrophobicity of the tip permits water to displace the resin from the tip (small or absent slope and/or selectivity and low electrode resistance). Controlled dry tip breakage on paper coated with glassine to final tip sizes well below 0.5 micron (confirmed by scanning electron microscopy) improves the yield of usable electrodes severalfold. Adequate silanization of the tip and consequent retention of resin at the tip can be predicted from the contact angles observed at the glass-LIE-backfilling solution interface. Satisfactory silanization can be achieved despite high ambient humidity. No evidence of shunting of Na+-LIE microelectrodes by the glass wall was seen. In the isolated perfused proximal tubule of Ambystoma tigrinum, the mean intracellular Na+ activity recorded by broken-tip electrodes (13.7 +/- 1.9 meq, n = 4) was similar to that recorded by intact electrodes (15.5 +/- 1.1 meq, n = 31).