Paper-based microfluidic devices exhibit many advantages for biological assays. Normally, the assays are restricted to certain areas of the paper by hydrophobic barriers comprised of wax or alkyl ketene dimers (AKD). Neither hydrophobic barrier is able to constrain aqueous solutions of surfactants, which are frequently used in biological assays. We demonstrate that rapidly curing silicone resins can be inkjet printed onto pure cellulose paper using inexpensive thermal ink-jet printers. The Piers-Rubinsztajn (PR) reaction dominates the cure chemistry leading to cellulose fibers that are surface coated with a silicone resin. The resulting barriers are able to resist penetration by surfactant solutions and even by the lower surface energy solvents DMF and DMSO. The utility of the barrier was demonstrated using a coliform assay based on detection of β-galactosidase.