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 N,N-dimethyl acrylamide and a benzophenone-based cross-linker,
which are photochemically transformed into a fiber-attached polymer
hydrogel on the paper fiber surfaces in situ. We show that the combination
of fluorescence detection and the use of a protein-repellent coating
enables sensitive paper-based analysis. Finally, the success of the
strategy is demonstrated by using a simple LFD application as an example.