Development of microfluidic impedance cytometry enabling the quantification of specific membrane capacitance and cytoplasm conductivity from 100,000 single cells.

This paper presents a new microfluidic impedance cytometry with crossing constriction microchannels, enabling the characterization of cellular electrical markers (e.g., specific membrane capacitance (Csm) and cytoplasm conductivity (σcy)) in large cell populations (~ 100,000 cells) at a rate greater than 100 cells/s. Single cells were aspirated continuously through the major constriction channel with a proper sealing of the side constriction channel. An equivalent circuit model was developed and the measured impedance values were translated to Csm and σcy. Neural network was used to classify different cell populations where classification success rates were calculated. To evaluate the developed technique, different tumour cell lines, and the effects of epithelial-mesenchymal transitions on tumour cells were examined. Significant differences in both Csm and σcy were found for H1299 and HeLa cell lines with a classification success rate of 90.9% in combination of the two parameters. Meanwhile, tumour cells A549 showed significant decreases in both Csm and σcy after epithelial-mesenchymal transitions with a classification success rate of 76.5%. As a high-throughput microfluidic impedance cytometry, this technique can add a new marker-free dimension to flow cytometry in single-cell analysis.