The changes in interparticle spacing upon hydration and dehydration of drop-cast films of hydrophilic gold nanoparticles (GNP) have been measured in situ with nanometer resolution using WetSTEM and ESEM. These subtle variations correlate well with the corresponding changes in the optical spectra and perceived color as well as changes in the electrical conductivity of the films. AC impedance analysis allows us to differentiate between resistive and capacitive components and to evaluate how these depend on average particle spacing and the water content of the matrix, respectively. Thin films of this type are well-known structures used for development of sensors and diagnostics.