Encyclopedia of Experiments: Biology
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Transcript
For the continuous measurement of cell behavior using real-time cell analysis, RTCA, obtain an RTCA microtiter plate containing impedance measuring-gold microelectrode arrays embedded in the well bottom. Carefully add cell culture medium to the wells.
Within the electrically conductive medium, apply a weak electrical potential across the microelectrodes. The potential causes electric current to flow between the microelectrodes, facilitating the measurement of the baseline impedance — the medium's inherent electrical resistance.
Pipette the desired concentrations of mammalian cell suspension to the medium-containing wells. Allow the cells to uniformly distribute in the well.
During incubation, the cells adhere onto the well bottom. The adherent cells on the microelectrodes act as an insulator, restricting the current flow and increasing the impedance.
Gradually, the cells proliferate on the microelectrodes, leading to increased exponential impedance.
Following confluent cell layer formation, the cells encounter nutrient deprivation and undergo death. As a result, the cells detach from the microelectrodes, decreasing the impedance.
The continuous impedance measurement facilitates real-time monitoring of the cellular events, including cell proliferation and death.
