Easily Fabricated Microfluidic Devices Using Permanent Marker Inks for Enzyme Assays

In this communication, we describe microfluidic paper analytical devices (μPADs) easily fabricated from commercially available Sharpie ink permanent markers on chromatography paper to colorimetrically detect glucose using glucose oxidase (GOx). Here, solutions of horseradish peroxidase (HRP), GOx, and potassium iodide (KI)were directly spotted onto the center of the μPAD and flowed into samples of glucose that were separately spotted on the μPAD. Using an XY plotter (Roland DGA Corporation, Irvine, CA USA), several ink marks drawn in the paper act as the hydrophobic barriers, thereby, defining the hydrophilic fluid flow paths of the solutions. Two paper devices are described that act as independent assay zones. The glucose assay is based on the enzymatic oxidation of iodide to iodine whereby a color change from clear to brownish-yellow is associated with the presence of glucose. In these experiments, two designs are highlighted that consist of circular paper test regions fabricated for colorimetric and subsequent quantification detection of glucose. The use of permanent markers for paper patterning is inexpensive and rapid and does not require special laboratory equipment or technical skill.