ECIS-Based Multi-Well Array Impedance Biosensor for the Comparison of Apoptosis Responses with Various Drug Testing

ECIS (Electric Cell-Substrate Impedance Sensing) based biosensors monitor physiological changes in cells by measuring electrical impedance in real-time, and have been actively utilized in various recent studies [3]. This non-invasive method requires no preprocessing and minimizes cell damage while providing highly sensitive and accurate data. The ECIS-based multi-well array impedance biosensor can analyze multiple samples simultaneously, making it suitable for high-throughput screening. Previous studies have developed and validated the reliability of this multi-well array impedance biosensor. The sensor features a 2x2 array pattern with independent environments, overcoming the limitation of traditional single-sensor single-sample measurements [1]. Additionally, the array impedance pattern allows the study of various drug responses at different concentrations and can be mass-produced at low cost through semiconductor processes.<br/> In this study, four anti-biotic drugs, Cycloheximide, Actinomycin D, Zeocin, and G418, which cause apoptosis and programmed cell death in NIH/3T3 cells, were monitor the cell-drug reaction for each drug, respectively. To prove the reliability of the sensor, Puromycin, a type of protein synthesis inhibitor, was used to induce cell death, which was accurately reflected through the impedance signal [2]. Based on this results, we conducted experiments on Cycloheximide, Actinomycin D, Zeocin, and G418, which have different biological mechanisms of action on NIH/3T3 cells. Cycloheximide blocks translation elongation by interfering with the movement of both tRNA and mRNA associated with the ribosome, Actinomycin D binds to DNA and blocks the production of RNA, Zeocin binds to DNA and induces double-strand breaks in DNA, and G418 binds to a subunit of the ribosome and stops translation of mRNA to protein, and in this way each drug induces cell death. We first defined a baseline of drug concentrations to be administered for comparative analysis with existing data for each drug. After setting the baseline concentration of each drug, the experiment was conducted by administering four concentrations with a certain ratio from the baseline concentration, and as a result, the cell death rate of different concentrations was accurately reflected through the impedance signal. In addition, the impedance signal accurately reflected the mechanism by which each drug induces cell death and the difference in cell behavior and death rate depending on the concentration, and the difference in cell death rate of each drug was quantitatively confirmed through mathematical analysis. Compared to JuLi Br, a real-time cell monitoring device, each impedance data accurately reflected the actual cell death behavior. Thus, the multi-well impedance biosensor accurately reflected the differences in cell behavior by concentration of four drugs with different biological mechanisms, which shows promise as a new drug development platform through drug compatibility evaluation to screen drugs and concentrations suitable for cancer patients in the future.<br/>References<br/>[1] D. H. Kang, et al, NPSM. 73,840-850 (2023)<br/>[2] G. Y. Lee, et al, Micromachines.12(10) 1248, (2021)<br/>[3] S.Arman, et al, Analyst. 149, 269-289, (2024)