Optical spectroscopy has been utilized in various fields of science, industry and medicine, since each substance is discernible from all others by its spectral properties. However, optical spectroscopy traditionally generates information on the bulk properties of the whole sample, and in applications in the agri-food industry for example some product properties result from the heterogeneity in its composition. This monitoring is considerably more challenging and can be successfully achieved by the so-called hyperspectral imaging technology, which allows the simultaneous determination of the optical spectrum and the spatial location of an object in a surface. In addition, it is a non-intrusive and non-contact technique which gives rise to a great potential for industrial applications and it does not require any particular preparation of the samples, which is a primary concern in food monitoring. This work illustrates an overview of approaches based on this technology to address different problems in agri-food, industrial and life-sciences sectors. The reviewed contributions comprise the doctoral thesis of Pilar Beatriz Garcia Allende and are focused on the development of new interpretation techniques and systems based on imaging spectroscopy. A hyperspectral imaging system was originally designed and tested for raw material on-line discrimination, which is a key factor in the input stages of many industrial sectors. The combination of the acquisition of the spectral information across transversal lines while materials are being transported on a conveyor belt, and appropriate image analyses have been successfully validated in the tobacco industry. Secondly, the use of imaging spectroscopy applied to online welding quality monitoring is discussed and compared with traditional spectroscopic approaches in this regard. Lastly, by means of the interpretation of light scattering signals that occur in biological tissue, a surgical guidance system to delineate tumor margins was developed.