Dielectrophoresis (DEP) manipulation combined with micro-electric impedance spectroscopy (µEIS) presents a sophisticated approach for cellular analysis and dielectric characterization. While conventional cell analysis techniques rely on complex labeling methods with inherent limitations, integrating DEP and µEIS offers non-invasive, label-free cellular characterization with enhanced sensitivity. This study presents an innovative dual-mode DEP platform incorporating both levitation (LEVDEP) and rotational (ROTDEP) forces, integrated with high-precision impedance measurement capabilities on one chip, enabling simultaneous Cell controlling and manipulation and dielectric signature extraction within a single microfluidic device. The fabricated and developed microfluidic platform demonstrated exceptional particle discrimination through the dual mode, with distinct responses for both particle populations. Under (F_{lEV.DEP}^{10.4 mu m}) 2.01 MHz showed a 63.4% magnitude increase, while (F_{lEV.DEP}^{24.9 mu m }) , particles exhibited a higher 81.2% increase at the same force, yielding a 2.48 × enhancement in discrimination ratio compared to no-DEP conditions. ROTDEP at 110 kHz induced even more pronounced differences, with (F_{ROT.DEP}^{10.4 mu m}) showing a 120% magnitude increase (phase patterns: −24.501° to −34.363°) and (F_{ROT.DEP}^{24.9 mu m}) µm particles demonstrating a 145% increase (phase patterns: −31.267° to −42.891°), achieving a 3.16 × discrimination ratio enhancement. The impedance spectrum revealed distinct frequency-dependent signatures, with ROTDEP showing superior mid-frequency discrimination (10.4 µm: 1.9370×({10}^{4}) Ω vs 24.9 µm: 2.0542×({10}^{4}) Ω at 110 kHz) and LEVDEP optimizing high-frequency characterization (10.4 µm: 1.6677×({10}^{4}) Ω vs 24.9 µm: 1.5849×({10}^{4}) Ω at 2.01 MHz). These signatures demonstrate the platform’s comprehensive particle characterization capabilities through complementary DEP forces. The dual-mode approach enhanced discrimination ratios by 2.48 × under (Lev. force) and 3.16 × under (LEV. force) at selected characteristic frequency range compared to (NonDEP force) conditions. Comprehensive impedance analysis through frequency spectrum (10 kHz—2.01 MHz) revealed unique frequency-dependent cell signatures, (ROT. force) demonstrating superior mid-frequency discrimination (magnitude differences of 1.9370 × 104 Ω vs 2.0542 × 104 Ω at 110 kHz) and LEVDEP optimizing high-frequency characterization (1.6677 × 104 Ω vs 1.5849 × 104 Ω at 2.01 MHz). Impedance dielectric analysis conducted over the 10 kHz to 2.01 MHz frequency range demonstrated frequency-dependent characteristics for each selected cell population. ROTDEP enhanced