Cellular subpopulations identified using an ensemble average of multiple dielectrophoresis measurements.

While the average value measurement approach can successfully analyze and predict the general behavior and biophysical properties of an isogenic cell population, it fails when significant differences among individual cells are generated in the population by intracellular changes such as the cell cycle, or different cellular responses to certain stimuli. Detecting such single-cell differences in a cell population has remained elusive. Here, we describe an easy-to-implement and generalizable platform that measures the dielectrophoretic cross-over frequency of individual cells by decreasing measurement noise with a stochastic method and computing ensemble average statistics. This platform enables multiple, real-time, label-free detection of individual cells with significant dielectric variations over time within an isogenic cell population. Using a stochastic method in combination with the platform, we distinguished cell subpopulations from a mixture of drug-untreated and -treated isogenic cells. Furthermore, we demonstrate that our platform can identify drug-treated isogenic cells with different recovery rates.

On-Chip Single-Cell Bioelectrical Analysis for Identification of Cell Electrical Phenotyping in Response to Sequential Electric Signal Modulation.

In recent years, an interesting biomarker called membrane breakdown voltage has been examined using artificial planar lipid bilayers. Even though they have great potential to identify cell electrical phenotyping for distinguishing similar cell lines or cells under different physiological conditions, the biomarker has not been evaluated in the context of living cell electrical phenotyping. Herein, we present a single-cell analysis platform to continuously measure the electric response in a large number of cells in parallel using electric frequency and voltage variables. Using this platform, we measured the direction of cell displacement and transparent cell image alteration as electric polarization of the cell responds to signal modulation, extracting the dielectrophoretic crossover frequency and membrane breakdown voltage for each cell, and utilizing the measurement results in the same spatiotemporal environment. We developed paired parameters using the dielectrophoretic crossover frequency and membrane breakdown voltage for each cell and evaluated the paired parameter efficiency concerning the identification of two different breast cancer cells and cell drug response. Moreover, we showed that the platform was able to identify cell electrical phenotyping, which was generated by subtle changes in cholesterol depletion-induced cell membrane integrity disruption when the paired parameter was used. Our platform introduced in this paper is extremely useful for facilitating more accurate and efficient evaluation of cell electrical phenotyping in a variety of applications, such as cell biology and drug discovery.

Ultra-sensitive dielectrophoretic surface charge multiplex detection inside a micro-dielectrophoretic device.

Label-free dielectrophoretic force-based surface charge detection has shown great potential for highly sensitive and selective sensing of metal ions and small biomolecules. However, this method suffers from a complex calibration process and measurement signal interference in simultaneous multi-analyte detection, thus creating difficulties in multiplex detection. We have developed a method to overcome these issues based on the optical discrimination of the dielectrophoretic behaviors of multiple microparticle probes considering the surface charge difference before and after self-assembling conjugation. In this report, we demonstrate and characterize this dielectrophoretic force-based surface charge detection method with particle probes functionalized by various biomolecules. This technique achieved an attomolar limit of detection (LOD) for Hg2+ in distilled water and a femtomolar LOD in drinking water using DNA aptamer-functionalized particle probes. More importantly, using two different DNA aptamer-functionalized particle probes for Hg2+ and Ag+, label-free dielectrophoretic multiplex detection of these species in drinking water with a femtomolar and a nanomolar LOD was achieved for the first time.