Traditional microfluidic paper-based analytical devices (μPADs) consist of a flat straight channel printed on a paper substrate. Such devices provide a promising low-cost solution for a variety of biomedical assays. However, they have a relatively high sample consumption due to their use of external reservoirs. Moreover, in μPADs based on the ion concentration polarization (ICP) effect, controlling the cross-sectional area of the Nafion membrane relative to that of the hydrophilic channel is difficult. Accordingly, the present study utilizes an origami technique to create a μPAD with a three-dimensional (3D) structure. The μPAD features short channels and embedded reservoirs, and therefore reduces both the driving voltage requirement and the sample consumption. Moreover, the preconcentration effect is enhanced through the use of an additional hydrophilic area adjacent to the Nafion membrane. The existence of electroosmotic flow (EOF) within the proposed device is confirmed using a current-monitoring method. In addition, the occurrence of ICP is evaluated by measuring the current–voltage response of the device at external voltages ranging from 0 to 50 V. The experimental results obtained for a fluorescein sample with an initial concentration of 10−5 M show that a 100-fold enhancement factor can be achieved given the use of a non-uniform-geometry design for the assay channel and an additional hydrophilic region with an area equal to approximately 10% of the channel cross-sectional area. Finally, a 100-fold factor can also be achieved for a fluorescein isothiocyanate sample with an initial concentration of 10−6 M given an external driving voltage of 40 V.
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