Micro-agar salt bridge in patch-clamp electrode holder stabilizes electrode potentials.
Maintaining a stable electrode potential is critical for patch-clamp measurements. The electrode potential of conventional patch electrode-holder assembly, where an Ag/AgCl wire is in direct contact with the patch pipette filling solution, is subject to drift if the pipette solution contains a low concentration of chloride ions (Cl-). We developed an agar bridge of 3 M KCl filled in a polyimide microtubing which forms an electrical connection between an Ag/AgCl wire and the pipette solution. We examined the offset potentials of the micro-agar salt bridge electrode assembly in parallel with a conventional electrode assembly in generic recording conditions (the pipette solution contained 5 mM NaCl). The junction potential between the Ag/AgCl wire and the pipette filling solution in the conventional electrode contributed to most of the offset potential drift observed during the course of 30 min recordings. The drift was up to 27.3 mV after several changes of the glass pipette. In contrast, the micro-agar salt bridge stabilized the electrode potential within typically 2 mV without affecting the patch electrode resistance, capacitance or noise level. Numerical simulations showed that Cl- diffusion from the agar bridge to the tip caused a negligible 0.4 microM Cl- concentration change at the pipette tip within 30 min. This method is easy to implement and provides long-term recording stability. The micro-agar salt bridge can fit in most commercial patch electrode holders and can be conveniently maintained.
Shao
Xuesi M
XM
Department of Neurobiology, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095-1763, USA. mshao@ucla.edu
Feldman
Jack L
JL
D000362:Agar; D000465:Algorithms; D002712:Chlorides / Q000737:chemistry; D003198:Computer Simulation; D003627:Data Interpretation, Statistical; D004058:Diffusion; D025802:Electric Capacitance; D017097:Electric Impedance; D004566:Electrodes; D008564:Membrane Potentials / Q000502:physiology; D036103:Nanotechnology; D018408:Patch-Clamp Techniques / Q000295:instrumentation*; D011189:Potassium Chloride / Q000737:chemistry
D002712:Chlorides; D011189:Potassium Chloride; D000362:Agar