Label-Free Multifrequency Impedance Detection System for Single-Cell Analysis: A Novel Approach for Hybridoma and Myeloma Cell Differentiation

Hybridoma cell screening is crucial for the production of monoclonal antibodies; however, traditional screening methods are often time-consuming and reliant on antigenantibody labeling. This study introduces a label-free, single-cell, multifrequency impedance detection system that innovatively utilizes impedance spectra as a novel physical parameter to differentiate between various hybridoma and myeloma cell types. The sensing device integrates dual-funnel structures with coplanar electrodes, which are, respectively, utilized for cell capture and impedance measurement. By optimizing the excitation voltage and flow rate, the system effectively reduces measurement errors. The sensing device was validated using polystyrene microbeads of different diameters, yielding the coefficients of variation (CV) of 0.0241%0.0132%, indicating high measurement stability and repeatability. The system was further validated with two types of hybridoma cells (E5, CD2) and myeloma cells (SP20), and impedance data were analyzed using orthogonal partial least squares discriminant analysis (OPLS-DA) via SIMCA 14.5 software, achieving a model prediction accuracy of 0.974. The results demonstrate that the system can stably and accurately measure cell impedance spectra and that impedance spectra, as a label-free parameter, can effectively distinguish between different types of hybridoma and myeloma cells, showing great potential for hybridoma cell sorting.