A Numerical Investigation of Enhancing Microfluidic Heterogeneous Immunoassay on Bipolar Electrodes Driven by Induced-Charge Electroosmosis in Rotating Electric Fields

A unique approach is proposed to boost on-chip immuno-sensors, for instance, immunoassays, wherein an antibody immobilized on the walls of a microfluidic channel binds specifically to an antigen suspended freely within a working fluid. The performance of these sensors can be limited in both susceptibility and response speed by the slow diffusive mass transfer of the analyte to the binding surface. Under appropriate conditions, the binding reaction of these heterogeneous immuno-assays may be enhanced by electroconvective stirring driven by external AC electric fields to accelerate the translating motion of antigens towards immobilized antibodies. To be specific, the phenomenon of induced-charge electroosmosis in a rotating electric field (ROT-ICEO) is fully utilized to stir analyte in the vicinity of the functionalized surface of an ideally polarizable floating electrode in all directions inside a tri-dimensional space. ROT-ICEO appears as a consequence of the action of a circularly-polarized traveling wave signal on its own induced rotary Debye screening charge within a bipolar induced double layer formed on the central floating electrode, and thereby the pertinent electrokinetic streamlines exhibit a radially converging pattern that greatly facilitates the convective transport of receptor towards the ligand. Numerical simulations indicate that ROT-ICEO can enhance the antigen–antibody binding reaction more effectively than convectional nonlinear electroosmosis driven by standing wave AC signals. The effectiveness of ROT-ICEO micro-stirring is strongly dependent on the Damkohler number as well as the Peclet number if the antigens are carried by a continuous base flow. Our results provide a promising way for achieving a highly efficient heterogeneous immunoassay in modern micro-total-analytical systems.

AC Electroosmosis Effect on Microfluidic Heterogeneous Immunoassay Efficiency

A heterogeneous immunoassay is an efficient biomedical test. It aims to detect the presence of an analyte or to measure its concentration. It has many applications, such as manipulating particles and separating cancer cells from blood. The enhanced performance of immunosensors comes down to capturing more antigens with greater efficiency by antibodies in a short time. In this work, we report an efficient investigation of the effects of alternating current (AC) electrokinetic forces such as AC electroosmosis (ACEO), which arise when the fluid absorbs energy from an applied electric field, on the kinetics of the antigen–antibody binding in a flow system. The force can produce swirling structures in the fluid and, thus, improve the transport of the analyte toward the reaction surface of the immunosensor device. A numerical simulation is adequate for this purpose and may provide valuable information. The convection–diffusion phenomenon is coupled with the first-order Langmuir model. The governing equations are solved using the finite element method (FEM). The impact of AC electroosmosis on the binding reaction kinetics, the fluid flow stream modification, the analyte concentration diffusion, and the detection time of the biosensor under AC electroosmosis are analyzed.

Heterogeneous Immunoassay Using Channels and Droplets in a Digital Microfluidic Platform

This work presents a heterogeneous immunoassay using the integrated functionalities of a channel and droplets in a digital microfluidic (DMF) platform. Droplet functionality in DMF allows for the programmable manipulation of discrete sample and reagent droplets in the range of nanoliters. Pressure-driven channels become advantageous over droplets when sample must be washed, as the supernatant can be thoroughly removed in a convenient and rapid manner while the sample is immobilized. Herein, we demonstrate a magnetic bead-based, enzyme-linked immunosorbent assay (ELISA) using ~60 nL of human interleukin-6 (IL-6) sample. The wash buffer was introduced in the form of a wall-less virtual electrowetting channel by a syringe pump at the flow rate of 10 μL/min with ~100% bead retention rate. Critical parameters such as sample wash flow rate and bead retention rate were optimized for reliable assay results. A colorimetric readout was analyzed in the International Commission on Illumination (CIE) color space without the need for costly equipment. The concepts presented in this work are potentially applicable in rapid neonatal disease screening using a finger prick blood sample in a DMF platform.

Investigation of Issues for the Accurate and Precise Measurement of an Analyte Using Surface-Enhanced Raman Scattering (SERS)

This paper builds on an earlier examination of the influence of sampling size and analyte surface density on the accuracy and precision of measurements using surface-enhanced Raman scattering (SERS) to read out heterogeneous immunoassays. Quantitation using SERS typically relies on interrogating a small area on the sample surface by using a micrometer-sized laser spot for signal generation. The information obtained using such a small portion of sample is then projected as being representative of the much larger sample, which can compromise the accuracy and precision of the measurement due to undersampling. For a heterogeneous immunoassay interrogated by SERS, quantitation is, therefore, sensitive to the size of the analyzed area and the surface density of the measured analyte. To identify conditions in which sampling error poses a threat to accuracy and precision, a simulation of a SERS immunoassay was developed and compared to experimental results. The simulation randomly distributes adsorbates across the capture surface and then measures the density of adsorbates inside areas of analysis of different sizes. This approach mimics the analysis of a heterogeneous immunoassay when using a Raman microscope with different laser spot sizes. The results of the simulations, which were confirmed experimentally by comparison to an immunoassay of human immunoglobulin G (IgG) show that the accuracy and precision of the measurement improved with larger analysis areas and higher analyte concentrations due to the increased apparent homogeneity of the analyte within the area of analysis. By imposing a threshold on precision (5%), we also begin to establish a framework for the parameters necessary to achieve reliable quantitative measurements (e.g., laser spot size, analyte concentration, and sample volume).