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The
ECIS core technology is based on a technique of measuring the change in
impedance of a small electrode to AC current flow. The heart of the
measurement is a specialized slide that has 8 individual wells for cell
culturing. The base of the device has an array of gold film electrodes
that connect to the ECIS electronics to each of the 8 wells.
8
Well Electrode Array |
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Array with the wells removed showing more clearly the patterns of gold and
insulating films
The current flows between a 250 µm diameter electrode and a larger counter
electrode using normal culture medium as the electrolyte.
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Electrode top view indicating AC current flow between the small active
electrode and the counter electrode
Without cells, the current flows unrestrained from the surface of the
electrodes.
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Electrode cross section
With
cells attached and spread upon this region, the current must now flow in
the spaces under and between the cells, as the cell membrane are
essentially insulators.
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This
is a micrograph of the 250 µm diameter gold electrode. The yellow area
outside the circular electrode is an insulating film that defines the
electrode perimeter. Both regions are excellent substrates for cell culture
and essentially mimic the surfaces of normal tissue culture ware.
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Shown are some fixed and stained NIH 3T3 cells.
At the start of the measurement the electrode has no cells attached to it
and the resistance is about 2000 ohms. Upon inoculation, cells anchor and
spread on the base of the well including the active 250 µm electrode. With
the presence of the cells, their insulating plasma membranes constrain the
electrical current and force it to flow in regions beneath and between the
cells. This convoluted current path causes large changes in the measured
impedance. Although this is taking place at both the small electrode as well
as at the counter electrode, the impedance of the small electrode is several
hundred times larger, and so the contribution of the large counter electrode
is a fraction of a percent and can be ignored. In addition to the overall
increase in the impedance, small fluctuations can be easily observed,
because the live cells continuously alter their morphology and hence the
impedance. With the confluent cell layer in place, the resistance now has
reached nearly 15,000 ohms. It is important to note that the AC current used
in making these measurements (approximately 1 microampere) and the resulting
voltage drops across the cells (a few millivolts) has no detectable effects
upon them; the measurement is non-invasive.
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This
technique is a patented technology known as ECIS, an acronym for
Electric Cell-substrate Impedance Sensing. Cell densities ranging from a
heavy confluent layer to very sparse layers can be measured with this
approach. The size of the electrodes restricts the maximum number of
anchored cells that can be observed from 100 to 1000 cells (dependent upon
the type of electrode array used in the instrument), but even a single
isolated cell results in impedance changes that can be monitored
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