Tailoring band alignment layer-by-layer using heterojunctions of two-dimensional (2D) semiconductors is an attractive prospect for producing next-generation electronic and optoelectronic devices that are ultra-thin, flexible, and efficient. 2D layers of transition metal dichalcogenides (TMDs) in laboratory devices have already shown favorable characteristics for electronic and optoelectronic applications. Despite these strides, a systematic understanding of how band alignment evolves from monolayer to multilayer structures is still lacking in experimental studies, which hinders development of novel devices based on TMDs. Here we 2 report on the local band alignment of monolayer, bilayer, and tri-layer MoS 2 on a 285-nm-thick SiO 2 substrate using a new approach to probe the occupied electronic states based on photoemission electron microscopy and deep ultraviolet light. Local measurements of the vacuum level and the valence band edge at the Brillouin zone center show that the addition of layers to monolayer MoS 2 increases the relative work function, and pushes the valence band edge toward the vacuum level. We also deduced n-type doping of few-layer MoS 2 and type-I band alignment across monolayer-to-bilayer and bilayer-to-trilayer lateral junctions. Conducted in isolation from environmental effects owing to the vacuum condition of the measurement and an insulating SiO 2 substrate, this study shows a new metrology to uncover electronic properties intrinsic to MoS 2 semiconducting layers and emerging 2D crystals alike.