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Comments and Recommendations
Membrane Damage
Recording from the same cell at a later time requires that the cell remain healthy during
the interim incubation. The less damage done to the membrane during handling and
impaling the cell, the happier it will be. Use of the BUZZ function should help minimize
the trauma from electrode penetration. Membrane damage can be further reduced by
properly isolating the experimental platform from vibration. Finally, hydraulically driven
micro-manipulators will also reduce membrane damage while the electrodes are
in the cell.
Repeated Recordings
Most recording sessions will involve repeating the above steps several times with many
different cells. Unless there is a concern about contamination of the bath solution
by something carried over from previous experiments, the pipettes can also be used
repeatedly. They should be free of debris and should have approximately the same
resistance as they had in the previous recording. A significantly higher resistance could
indicate that the pipette is partially plugged with cellular debris.
Make the following control settings before the next recording is carried out:
Control
polarity
mode select
gain
Electrophysiology
If you are well versed in setting up electrophysiological equipment, you can safely skip
over the rest of this section. If, however, this is your initiation into electrophysiology (as
it may well be for some of you molecular biologists), then you may find the following
recommendations helpful.
While the whole cell electrophysiological configuration is more forgiving than the
patch clamp, it is still important to minimize mechanical motion. The platform for your
experimental setup, therefore, should be mechanically well isolated. This will reduce
leakage around the electrodes, making the clamp more effective and reducing noise in
your recordings. The latter is especially important when recording responses of certain
ligand-gated channels where membrane potential changes may only be a few mV
32
Section
command
clamp
clamp
Publication 5720-001 REV1.0
Setting
off
off
0 (fully CCW)
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