The biogeochemical functioning of soils (e.g., soil carbon stabilization and nutrient cycling) is determined at the interfaces of specific soil structures (e.g., aggregates, particulate organic matter (POM) and organo-mineral associations). With the growing accessibility of spectromicroscopic techniques, there is an increase in nano- to microscale analyses of biogeochemical interfaces at the process scale, reaching from the distribution of elements and isotopes to the localization of microorganisms. A widely used approach to study intact soil structures is the fixation and embedding of intact soil samples in resin and the subsequent analyses of soil cross-sections using spectromicroscopic techniques. However, it is still challenging to link such microscale approaches to larger scales at which normally bulk soil analyses are conducted. Here we report on the use of laboratory imaging Vis–NIR spectroscopy on resin embedded soil sections and a procedure for supervised image classification to determine the microscale soil structure arrangement, including the quantification of soil organic matter fractions. This approach will help to upscale from microscale spectromicroscopic techniques to the centimetre and possibly pedon scale. Thus, we demonstrate a new approach to integrate microscale soil analyses into pedon-scale conceptual and experimental approaches. Highlights: Quantification of soil constituents using Vis-NIR spectroscopy. New approach to use resin embedded soil core sections with intact structure. Reproducible quantification of soil constituents important for soil carbon storage. Vis-NIR as promising tool for upscaling from microscale to pdeon scale.