A Numerical Investigation of Enhanced Microfluidic Immunoassay by Multiple-Frequency Alternating-Current Electrothermal Convection

Compared with traditional immunoassay methods, microfluidic immunoassay restricts the immune response in confined microchannels, significantly reducing sample consumption and improving reaction efficiency, making it worthy of widespread application. This paper proposes an exciting multi-frequency electrothermal flow (MET) technique by applying combined standing-wave and traveling-wave voltage signals with different oscillation frequencies to a three-period quadra-phase discrete electrode array, achieving rapid immunoreaction on functionalized electrode surfaces within straight microchannels, by virtue of horizontal pumping streamlines and transverse stirring vortices induced by nonlinear electrothermal convection. Under the approximation of a small temperature rise, a linear model describing the phenomenon of MET is derived. Although the time-averaged electrothermal volume force is a simple superposition of the electrostatic body force components at the two frequencies, the electro-thermal-flow field undergoes strong mutual coupling through the dual-component time-averaged Joule heat source term, further enhancing the intensity of Maxwell–Wagner smeared structural polarization and leading to mutual influence between the standing-wave electrothermal (SWET) and traveling-wave electrothermal (TWET) effects. Through thorough numerical simulation, the optimal working frequencies for SWET and TWET are determined, and the resulting synthetic MET flow field is directly utilized for microfluidic immunoassay. MET significantly promotes the binding kinetics on functionalized electrode surface by simultaneous global electrokinetic transport along channel length direction and local chaotic stirring of antigen samples near the reaction site, compared to the situation without flow activation. The MET investigated herein satisfies the requirements for early, rapid, and precise immunoassay of test samples on-site, showing great application prospects in remote areas with limited resources.