Submicron-precision particle characterization in microfluidic impedance cytometry with double differential electrodes

Submicron-precision particle characterization is crucial for counting, sizing and identifying a variety of biological particles, such as bacteria and apoptotic bodies. Microfluidic impedance cytometry has been attractive in current research for microparticle characterization due to its advantages of label-free detection, ease of miniaturization and affordability. However, conventional electrode configurations of three electrodes and floating electrodes have not yet demonstrated the capability of probing submicron particles or microparticles with a submicron size difference. In this study, we present a label-free high-throughput (∼800 particles per second) impedance-based microfluidic flow cytometry system integrated with a novel design of a double differential electrode configuration, enabling submicron particle detection (down to 0.4 μm) with a minimum size resolution of 200 nm. The signal-to-noise ratio has been boosted from 13.98 dB to 32.64 dB compared to a typical three-electrode configuration. With the proposed microfluidic impedance cytometry, we have shown results of sizing microparticles that accurately correlate with manufacturers' datasheets (R² = 0.99938). It also shows that population ratios of differently sized beads in mixture samples are consistent with the results given by commercial fluorescence-based flow cytometry (within ∼1% difference). This work provides a label-free approach with submicron precision for sizing and counting microscale and submicron particles, and a new avenue of designing electrode configurations with a feature of suppressing the electrical noise for accomplishing a high signal-to-noise ratio in a wide range of frequencies. This novel double differential impedance sensing system paves a new pathway for real-time analysis and accurate particle screening in pathological and pharmacological research.