A Microfluidic Flow Cytometer Composed of Double T-Type Constriction Channel with Predefined Fluorescence Detection Window, Enabling High-Throughput Characterization of Intrinsic Single-Cell Structural and Electrical Parameters

This paper presented a microfluidic flow cytometer composed of a double T-type constriction channel with a predefined fluorescence detection window, enabling high-throughput characterization of intrinsic single-cell structural parameters of cell diameter (Dc)/nuclear diameter (D n ), and electrical parameters of specific membrane capacitance (Csm)/cytoplasmic conductivity ( $\sigma_{\text{cy}}$ ) based on a home-developed equivalent impedance-fluorescence model. As a demonstration, D c , D n , C sm and $\sigma_{\text{cy}}$ of single blood cells were quantified as $15.18\pm 1.00$ vs. $13.99\pm 1.91\ \mu\mathrm{m},\ 11.83\pm 1.75$ vs. $10.59\pm 1.35\ \mu \mathrm{m},\ 2.78\pm 0.41$ vs. $2.84\pm 0.84\ \mu\mathrm{F}/\text{cm}^{2}$ and $0.86\pm 0.19$ vs. $0.74\pm 0.35\ \mathrm{S}/\mathrm{m}$ for K562 ( $\mathrm{N}_{\text{cell}}=2173$ ) vs. Jurkat ( $\mathrm{N}_{\text{cell}}=1326$ ) cells. In comparison to hematology analyzers capable of only quantifying intrinsic biophysical parameters of D c , the microfluidic flow cytometer developed in this study can quantify four intrinsic parameters of D c , D n , C sm and $\sigma_{\text{cy}}$ simultaneously and improve the cell-type classification from 68.7% (D c ) to 84.0% (D c , D n , C sm and $\sigma_{\text{cy}}$ ).