A substrate inspection apparatus is disclosed. The substrate inspection apparatus includes: a first light source configured to radiate an ultraviolet light onto a coated film of a substrate, the coated film being mixed with fluorescent pigments; a first light detector configured to capture fluorescence generated from the coated film onto which the ultraviolet light is radiated, and to obtain a two-dimensional (2D) image of the substrate; a processor configured to derive one region among a plurality of regions of the substrate based on the 2D image; a second light source configured to radiate a laser light onto the one region; and a second light detector configured to obtain optical interference data generated from the one region by the laser light, wherein the processor is configured to derive a thickness of the coated film of the one region based on the optical interference data.