Phenotypic quantification of cells based on their plasma membrane capacitance and cytoplasmic conductivity, as determined by their dielectrophoretic frequency dispersion, is often used as a marker for their biological function. However, due to the prevalence of phenotypic heterogeneity in many biological systems of interest, there is a need for methods capable of determining the dielectrophoretic dispersion of single cells at high throughput and without the need for sample dilution. We present a microfluidic device methodology wherein localized constrictions in the microchannel are used to enhance the field delivered by adjoining planar electrodes, so that the dielectrophoresis level and direction on flow-focused cells can be determined on each traversing cell in a high-throughput manner based on their deflected flow streamlines. Using a sample of human red blood cells diluted to 2.25 × 108 cells/mL, the dielectrophoretic translation of single cells traversing at a flow rate of 1.68 μL/min is measured at a throughput of 1.1 × 105 cells/min, to distinguish positive versus negative dielectrophoresis and determine their crossover frequency in media of differing conductivity for validation of the computed membrane capacitance to that from prior methods. We envision application of this dynamic dielectrophoresis (Dy-DEP) method towards high-throughput measurement of the dielectric dispersion of single cells to stratify phenotypic heterogeneity of a particular sample based on their DEP crossover frequency, without the need for significant sample dilution.
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We thank Dr. Jennifer Guler (PI of the Malaria Lab, University of Virginia) and graduate student Audrey Brown for providing the RBC samples used in this work.
Funding from NIH grants 1R21AI130902-01 and R01 CA200755, Advanced Regenerative Medicine Institute’s BioFab, USA, Subcontract T0163, and University of Virginia’s 3C program are acknowledged.
The reported studies on blood samples have been approved by the University of Virginia Institutional Review Board for Health Sciences Research (IRB-HSR protocol #21081) and have been performed in accordance with ethical standards.
Conflict of interest
The authors declare that they have no conflict of interest.
Published in the topical collection Bioanalytics and Higher Order Electrokinetics with guest editors Mark A. Hayes and Federica Caselli.
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Torres-Castro, K., Honrado, C., Varhue, W.B. et al. High-throughput dynamical analysis of dielectrophoretic frequency dispersion of single cells based on deflected flow streamlines. Anal Bioanal Chem 412, 3847–3857 (2020). https://doi.org/10.1007/s00216-020-02467-1