Controlling Fluorescent Readout in Paper-based Analytical Devices.

Paper is an ideal candidate for the development of new disposable diagnostic devices because it is a low-cost material, allows transport of the liquid on the device by capillary action, and is environmentally friendly. Today, colorimetric analysis is most often used as a detection method for rapid tests (test strips or lateral flow devices) but usually gives only qualitative results and is limited by a relatively high detection threshold. Here, we describe studies using fluorescence as a readout tool for paper-based diagnostics. We study how the optical readout is affected by light transmission, scattering, and fluorescence as a function of paper characteristics such as thickness (grammage), water content, autofluorescence, and paper type/composition. We show that paper-based fluorescence analysis allows better optical readout compared to that of nitrocellulose, which is currently the material of choice in colorimetric assays. To reduce the loss of analyte molecules (e.g., proteins) due to adsorption to the paper surface, we coat the paper fibers with a protein-repellent hydrogel. For this purpose, we use hydrophilic copolymers consisting of