Membrane electrophysiology practical exercise

Ag/AgCl electrodes in physiology

Ag/AgCl electrodes, often used in electrophysiology, consist of silver wire with a silver chloride coating
(the coating is grown on the wire by electrolysis in 1M HCl). The coating will ionise partly in solution.
The electrodes undergo the following reversible reaction with the solution surrounding them:

Ag (s) + Cl- (aq) ⇋ AgCl (s) + e-

When placed on either side of the artificial membrane in

our practical, across which is a voltage, silver loses an
electron at one electrode and forms AgCl with chloride
from the solution (the electrode at which this happens is
by definition the anode, which is technically an electrode
receiving positive current from an external circuit, i.e.
electrons are moving away from this electrode in the wire).
The electron travels through the wire and is then used to
convert a silver ion from AgCl into the metallic form at the
other electrode (by definition the cathode); a chloride ion
goes into solution. The potential difference between the
two sides is recorded by a voltmeter, in our case
PowerLab.

The problem is that the reaction above, and thus the

response of each Ag/AgCl electrode, is sensitive not just to the potential difference across the
membrane, but also to different chloride concentrations in the bulk solution: the electrodes will
generate different “junction potentials” as the chloride levels are changed. If the junction potential at
one electrode changes, the potential difference recorded by the voltmeter between the two electrodes
will vary. Changing chloride levels throughout an experiment using such electrodes could therefore
result in a voltage drift, which would contaminate whatever it was that you wanted to measure (the
voltage across the membrane, in our case). However, if the junction potential is the same at each
electrode and can be held constant throughout the experiment, the potential difference between the
electrodes resulting from this effect would be zero, and any voltage recorded would then be the
voltage that you are trying to measure.

The agar surrounding the electrodes in our experiment contains KCl at a fixed, very high
concentration (1-3 M), and is used as an electrolyte layer or “agar bridge”. Because it is ion-
permeable, the agar maintains electrical contact with the bulk solution while at the same time keeping
a constant chloride ion concentration in contact with the electrodes, which is not subject to change
when the test solution is changed. This makes use of the fact that agar eliminates convection
currents, and thus greatly reduces the mixing of the solution immediately surrounding the electrode
with the test solution added to the cell during the experiment (diffusion remains, but this is extremely
slow). Because this keeps the junction potentials on each side constant, they cancel each other out
and can therefore be ignored: what we record is the membrane potential only, which is what we want.

Copyright © Matthew J. Mason, 2020