Cellular Impedance-Based Analysis for Monitoring of Cellular Processes in Real-Time

Abstract

Source: Labadie, T., et al., Monitoring Influenza Virus Survival Outside the Host Using Real-Time Cell Analysis. J. Vis. Exp. (2021)

In this video, we demonstrate the process of real-time monitoring of cellular events, including the proliferation of adherent cells, via a cellular impedance assay. Here, the adherent cells act as insulators, restricting the current flow between the electrodes and increasing the impedance.

Protocol

NOTE: During all experiments, keep the plates on non-electrostatic surfaces at all times, such as the paper wraps from the packaging. Follow section 1 below to determine the most appropriate concentration of cells to be seeded in the electronic microtiter plate (designed as E-Plate).

1. Determination of appropriate cell quantity for infecting cells

1. Prepare MDCK (Madin-Darby Canine Kidney) cells in 75 cm² flasks to obtain freshly split cells (approximately 80% confluence) 24 h before the experiment.
2. Wash cells with 5 mL of 1x PBS and detach them by adding 3 mL of 0.25% Trypsin-EDTA solution.<....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
E-Plate 16 (6 plates)	ACEA Biosciences, Inc	5469830001	E-plates are available in different packaging
PBS 1X	Life technologies (gibco)	14040091
TPCK-Trypsin	Worthington	LS003740
xCELLigence Real-Time Cell Analysis Instrument S16	ACEA Biosciences, Inc	380601310	The xCELLigence RTCA S16 instruments are available in different formats (16-well, 96-well, single or multi-plate)
75 cm2 tissue culture flask	Falcon	430641U
MEM 1X	Life technologies (gibco)	31095029