Three-Dimensional Paper-Based Microfluidic Analytical Devices Integrated with a Plasma Separation Membrane for the Detection of Biomarkers in Whole Blood

Abstract

Paper-based microfluidic analytical devices (μPADs) have recently attracted attention as a point-of-care test kit because of their low cost and nonrequirement for external forces. To directly detect biomarkers in whole blood, however, they need to be assembled with a filter such as a plasma separation membrane (PSM) because the color of the blood cells interferes with the colorimetric assay. However, this assembly process is rather complicated and cumbersome, and the fluid does not uniformly move to the detection zone when the adhesion between the paper and PSM is not perfect. In this study, we report a simple three-dimensional (3D) printing method for fabricating PSM-integrated 3D-μPADs made of plastics without the need for additional assembly. In detail, PSM was coated with parylene C to prevent its dissolution from organic solvent during 3D printing. Then, the coated PSM was superimposed on the paper. Detection zones and a reservoir were printed on the paper and PSM via liquid photopolymerization, using a digital light processing printer. The limit of detection of the PSM-integrated 3D-μPADs for glucose in whole blood was 0.3 mM, and these devices demonstrated clinically relevant performance on diabetes patient blood samples. Our 3D-μPADs can also simultaneously detect multiple metabolic disease markers including glucose, cholesterol, and triglycerides in whole blood. Our results suggest that our printing method is useful for fabricating 3D-μPADs integrated with PSM for the direct detection of biomarkers in whole blood.

Three-Dimensional Paper-Based Microfluidic Analysis Device for Simultaneous Detection of Multiple Biomarkers with a Smartphone

“…The height of the structure cured by the resin in this study increased with rising exposure time. This is because the amount of cross-linked resin increases as the UV exposure time increases [ 10 , 20 ]. In fact, the reservoir at 5 mm height was printed by overexposing the paper for 60 s ( Figure 2 b).…”

“…This also reduces the amount of the sample, which leads to the detection of more biomarkers. In previous research, we succeeded in integrating plasma separation membrane with 3D-µPADs [ 20 ]. This suggests that the 3D-µPADs with the plasma separation membrane will allow patients to easily diagnose diseases remotely using a smartphone at home in the future.…”

“…Smartphones can provide easy detection and quick analysis results for POCT kits [ 16 , 17 , 18 ]. For example, smartphone-based colorimetric and fluorescence analyses have been extensively studied for medical diagnostics [ 19 , 20 ], environmental monitoring [ 21 ], and food evaluation [ 22 ]. So far, only a small number of biomarkers can be analyzed using a smartphone, making it difficult to accurately diagnose diseases.…”

Three-Dimensional Paper-Based Microfluidic Analysis Device for Simultaneous Detection of Multiple Biomarkers With a Smartphone

“…This is because the amount of cross-linked resin increases as the UV exposure time increases. [9,19] In fact, the reservoir at 5 mm height was printed by overexposing the paper for 60 s. It took ~90 s to print the top and bottom structures of the paper and remove uncured resin from the paper (Figure 2b). The results suggest that our new method is highly suitable for the rapid production of 3D-μPADs.…”

“…Smartphones can provide easy detection and quick analysis results for POCT kits [15][16][17]. For example, smartphone-based colorimetric and fluorescence analyses have been extensively studied for medical diagnostics [18,19], environmental monitoring [20], and food evaluation [21]. So far, only a small number of biomarkers can be analyzed using a smartphone, making it difficult to accurately diagnose diseases.…”