Micro-fabrication by wax spraying for rapid smartphone-based quantification of bio-markers

A cheap/rapid technique for the fabrication of μPADs is presented for point of care analysis. Hydrophobic channels were formed across the width of the paper by spraying hot pure wax. This biocompatible novel process yielded uniform 300 ± 5 μm hydrophilic microchannels on paper, without the use of the cleanroom, UV lamp, or organic solvents and was completed in a single step without the need for a hotplate. Hot wax is properly impregnated across paper thickness by spraying under optimized temperature and pressure. Our method is advantageous in the cost and ease of fabrication, process time (<1 min), the feasibility of mass-fabrication, readout, environmental considerations and multiplexing due to the embossed structure of remnant wax. The performance of the resulting μPAD was assessed on a multiplexed Uric acid and Nitride assay, bearing 95% of confidence level in the readout against standardized tests. A novel RGB processing app was developed for smart-phones to quantify colorimetric read-outs through a heuristic normalization equation that converts RGB to integer systems. This combinatorial sensor demonstrates a good linear range (up to 800 μM for Uric acid and 1250 μM for Nitride), low detection limit (100 μM for uric acid and 156 μM for nitride).