Abstract
An improved approach for comparative study of plant cells for long term and continuous monitoring using electrical impedance spectroscopy is demonstrated for tomato and tobacco plant cells (MSK8 and BY2) in suspensions. This approach is based on the locations and magnitudes of defining features in the impedance spectra of the recently reported unified equivalent circuit model. The ultra-wide range (4 Hz to 20 GHz) impedance spectra of the cell lines were measured using custom probes, and were analyzed using the unified equivalent circuit model, highlighting significant negative phase peaks in the ~ 1 kHz to ~ 10 MHz range. These peaks differ between the tomato and tobacco cells, and since they can be easily defined, they can potentially be used as the signal for differentiating between different cell cultures or monitoring them over time. These findings were further analysed, showing that ratios relating the resistances of the media and the resistance of the cells define the sensitivity of the method, thus affecting its selectivity. It was further shown that cell agglomeration is also an important factor in the impedance modeling in addition to the overall cell concentration. These results can be used for optimizing and calibrating electrical impedance spectroscopy-based sensors for long term monitoring of cell lines in suspension for a given specific cell and media types.
Highlights
An improved approach for comparative study of plant cells for long term and continuous monitoring using electrical impedance spectroscopy is demonstrated for tomato and tobacco plant cells (MSK8 and BY2) in suspensions
Tomato (S. lycopersicum cv Mill.; line M SK835) and tobacco (N. tabacum cv B Y236) cell suspension cultures were grown in Murashige and Skoog (MS)[37] media including vitamins (Duchefa Biochemie) in 250 mL flask with 100 mL liquid, supplemented with 30 g/L sucrose, 1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.1 mg/L kinetin, which was set to pH 5.7
Rs, Rct, Cdl, and the CPE as are defined in a standard Randles model, R represents the resistance of the cells, R1 represents the resistance of the solution, C represents the capacitance due to the presence of cells in the suspension, and C1 represents the capacitive effects of water polarization
Summary
An improved approach for comparative study of plant cells for long term and continuous monitoring using electrical impedance spectroscopy is demonstrated for tomato and tobacco plant cells (MSK8 and BY2) in suspensions This approach is based on the locations and magnitudes of defining features in the impedance spectra of the recently reported unified equivalent circuit model. The ultra-wide range (4 Hz to 20 GHz) impedance spectra of the cell lines were measured using custom probes, and were analyzed using the unified equivalent circuit model, highlighting significant negative phase peaks in the ~ 1 kHz to ~ 10 MHz range These peaks differ between the tomato and tobacco cells, and since they can be defined, they can potentially be used as the signal for differentiating between different cell cultures or monitoring them over time. The theoretical analyses are presented and discussed critically, and are applied to the experimentally obtained data in order to demonstrate their efficacy with regards to discrimination between cell and medium types
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