Abstract
<jats:p>Circulating tumor cells (CTCs) are valuable biomarkers for cancer diagnosis and monitoring, yet their isolation from blood remains challenging due to their phenotypic heterogeneity and rarity. Labelfree microfluidic technologies offer a promising alternative to affinity-based approaches by exploiting intrinsic biophysical differences between cell types. Here, we developed a microfluidic platform for label-free cell separation based on insulator-based dielectrophoresis (iDEP). The microfluidic device employs an array of triangular insulating structures that generate strong electric field gradients in response to an externally applied alternating current (AC) electric field, enabling selective isolation of breast cancer cells from blood cells based on their dielectric properties. Hydrodynamic focusing is used to confine the sample stream and precisely control cell trajectories within the separation region. Numerical simulations were performed to optimize the electric field distribution and fluid flow characteristics within the device. Experimental validation using breast cancer cell lines (mesenchymallike MDA-MB-231 cells and epithelial-like MCF-7 cells) spiked into peripheral blood mononuclear cells (PBMCs) demonstrates selective dielectrophoretic deflection of cancer cells while PBMCs largely follow the central streamline. The platform achieves recovery rates exceeding 98% and a separation purity above 65% within the optimized operating conditions. The proposed system provides a simple label-free approach to separate heterogeneous cell populations and represents a promising tool for microfluidic liquid biopsy enrichment applications.</jats:p>