Poly(ethylene glycol) (PEG) hydrogels have been used to encapsulate fluorescently labeled molecules in order to detect a variety of analytes. The hydrogels are designed with a mesh size that will retain the sensing elements while allowing for efficient diffusion of small analytes. Some sensing assays, however, require a conformational change or binding of large macromolecules, which may be sterically prohibited in a dense polymer matrix. A process of hydrogel microporation has been developed to create cavities within PEG microspheres to contain the assay components in solution. This arrangement provides improved motility for large sensing elements, while limiting leaching and increasing sensor lifetime. Three hydrogel compositions, 100% PEG, 50% PEG, and microporated 100% PEG, were used to create pH-sensitive microspheres that were tested for response time and stability. In order to assess motility, a second, more complex sensor, namely a FITC-dextran/TRITC-Con A glucose-specific assay was encapsulated within the microspheres.