Superhydrophobic Glass Microfiber Filter as Background-Free Substrate for Quantitative Fluorometric Assays

We have discovered herein that commonly used laboratory glass microfiber filters can be functionalized as background-free superhydrophobic substrates for quantitative fluorometric assays. In particular, glass microfiber filters (Whatman GF/A) can be treated with low-concentration (20 mM) methyltrichlorosilane/toluene solution to be superhydrophobic (water contact angle >150 degrees) in less than 5 mM; the modified glass microfiber filter can be readily patterned with UV/ozone irradiation to create hydrophilic reaction zones on the otherwise superhydrophobic substrate. Compared with traditional cellulose filter paper, the glass microfiber filter has extremely low fluorescence background, which makes it an excellent substrate for preparing quantitative fluorometric assays. In conjunction with smartphone imaging and color analysis, we have showcased a copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC)-based fluorometric assay for copper quantitation on these patterned, superhydrophobic glass microfiber filter substrates. Both the limit of detection and linear response range are comparable with the standard spectrophotometric quantitation in solution and commercial copper detection kits, which augments the application potential of superhydrophobic glass microfiber filters as ideal (e.g., background-free) substrates for the preparation of multiplex microassays and other advanced microanalytical devices based on fluorescence readout.