We have designed, fabricated, calibrated and tested actuators for shear characterization to assess microscale shear properties of soft substrates. Here we demonstrate characterization of dry silicone and hydrated polyethelyne glycol. Microscale tools, including atomic force microscopes and nanoindenters, often have limited functionality in hydrated environments. While electrostatic comb-drive actuators are particularly susceptible to moisture damage, through chemical vapor deposition of hexamethyldisiloxane, we increase the hydrophobicity of our electrostatic devices to a water contact angle 90 ± 3°. With this technique we determine the effective shear stiffness of both dry and hydrated samples for a range of soft substrates. Using computational and analytical models, we compare our empirically determined effective shear stiffness with existing characterization methods, rheology and nanoindentation, for samples with shear moduli ranging from 5-320 kPa. This work introduces a new approach for microscale assessment of synthetic materials that can be used on biological materials for basic and applied biomaterials research.