Cell-Based Detection Using Electric Cell-Impedance Sensing
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Cell-Based Detection Using Electric Cell-Impedance Sensing Bhavana Mohanraj1, Nathan Schiele1, Anne Hynes2 , Zijie Yan2 David T. Corr1, Cerasela Zoica Dinu3, Douglas B. Chrisey2 Rensselaer Polytechnic Institute (1) Biomedical Engineering, (2) Materials Science and Engineering, Troy, NY West Virginia University (3) Chemical Engineering, Morgantown, WV ABSTRACT Electric Cell-Impedance Sensing (ECIS) is a real-time transduction system that can be used to detect the presence of foreign particles or pathogens by measuring the changes in impedance or resistance of a cell monolayer grown on an electrode. Herein, we present the use of ECIS for the detection of the toxicity of silver nanoparticles on Madine Derby Canine Kidney (MDCK) epithelial cells as a function of changes in the cell confluence and barrier function of the cell monolayer. The barrier function is a measure of the number of tight junctions formed between confluent cells in a monolayer; tighter confluence leads to an increase in a barrier function and thus in the measured resistance. We were able to detect exposures as low as 1 µg of 20 nm silver nanoparticles per 105 cells within 2 hours; those exposures were quantified as a significant drop in impedance and a gradual decrease in the barrier function as compared to the controls. Future work would include the detection of protein toxins using impedance sensing as well as further analysis of the barrier function using fluorescent staining. INTRODUCTION Cell-based sensing systems aim to detect, in real-time, the presence of foreign pathogens or other agents which cause immediate changes in cell behavior that can be monitored using transduction mechanisms. One such system, Electric Cell-Impedance Sensing, or ECIS developed by Applied Biophysics, Troy, NY, applies an oscillating electric field to cells cultured on electrodes, and measures the resulting impedance as a function of frequency as the cells attach and spread on the electrode surface [1]. ECIS has the ability to detect cellular micro-motion as well as overall cellular morphological changes using electrodes of 250 µm in diameter, [2]. In previous work by Curtis et al., ECIS was used to quantify the toxicity of chemical agents such as mercury and arsenic that have potential applications in chemical warfare [4]. Other work also includes the use of ECIS to monitor the cytopathic effect due to infection of cultured cells by influenza virus A [2]. In our experiments, we explored the novel application of ECIS for the detection of nanoparticle toxicity in mammalian epithelial cells. Nanomaterials are a rapidly expanding field with potential applications in tissue engineering and materials development (10). However, the toxicity of these nanomaterials must be investigated prior to their use in new and advancing technologies. Previous studies have shown that silver nanoparticles for instance, have a strong cytotoxic effect on mouse germ and rat liver cells, although the mechanism of toxicity is not fully understood [5,6]. ECIS is a system which has
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