Abstract: A synthetic hydrogel is described, including hydrated mucin glycoproteins cross-linked with multi-arm thiol functional cross-linker, which can be prepared to model viscoelastic and micro-rheological properties of natural mucus. Such synthetic hydrogel can be prepared from a wide variety of mucin raw materials. Also described is a method of microrheologically characterizing mucus, by dispersing in the mucus muco-inert particles (MIP), irradiating the mucus containing MIP with polarized light, and measuring fluorescence polarization (FP) resulting from rotational diffusion of the MIP in the mucus in response to such irradiating, as a microrheological characteristic of the mucus. This method can be carried out using a plate reader equipped with a spectrofluorometer and polarized filter set, and therefore can be readily carried out in clinical settings without the necessity of specialized microrheological equipment.