Colloidal lithography is widely used as a low cost and large-area deposition approach, alternative to the conventional small-area expensive lithographic techniques, for the fabrication of short-range ordered sub-wavelength metallic nanostructures. This paper contributes to the understanding of the impact of the fabrication protocol of a colloidal mask on the optical and sensing properties of short range-ordered nanohole (NH) distributions fabricated by colloidal lithography in optically thin (20 nm thick) gold films. We consider polystyrene nanospheres (PS-NSPs) with a nominal diameter of 80 nm, electrostatically adsorbed from a salt-free colloidal solution onto a polydiallyldimethylammonium (PDDA) countercharged monolayer. By avoiding the conventional polyelectrolyte multilayer and based on the interplay between the deposition times of both PDDA and PS-NSPs, we demonstrate effective simplification of the commonly applied deposition protocol and effective tuning of the NH-to-NH spacing (dNN) with negligible agglomeration. Comparison with NH samples prepared by salt-containing colloidal solutions points out the negative impact of salt addition on the optical properties. The effective tuning of dNN obtained by our protocol demonstrates highly correlated disorder under unsaturated adsorption and allows a discussion on the analogies of the optical response between long- and short- range ordered NH systems, which is a still debated topic. By Fast Fourier Transform of autocorrelation images of scanning electron microscopy micrographs we demonstrate quantitatively, rather than in principle, the correspondence between an inherent ordering length-scale and dNN. As optical transducers for detecting refractive index changes, our samples exhibit significant bulk sensitivity (∼309 nm RIU-1) in the framework of short range ordered NH systems.