Abstract: An enhanced single particle interferometric reflectance imaging system includes an illumination source configured to produce illumination light along an illumination path toward a target substrate. The target substrate can be configured to reflect the illuminating light along one or more collection paths toward one or more imaging sensors. The target substrate includes a base substrate having a first reflecting surface and a transparent spacer layer having a first surface in contact with the first reflecting surface and a second reflecting surface on a side opposite to the first surface. The transparent spacer layer has a predefined thickness that is determined as a function of a wavelength of the illuminating light and produces a predefined radiation pattern of optical scattering when nanoparticles are positioned on or near the second reflective surface. In addition, one or more of the collection paths can also include an amplitude mask selected to match the radiation pattern.