Wheat bran comprises an underutilized resource. As a byproduct of white flour production, it is primarily used as animal feed. This work was conducted in the framework of a project that deals with the biorefinery of wheat bran, investigating advanced analytical strategies for in-depth characterization of wheat bran and wheat bran fractions. Three sets of tools were established: structural analysis of polysaccharides, visualization techniques, and rapid screening methods. Methylation analysis followed by methanolysis revealed the linkages between the monosaccharide components of arabinoxylan. It was applied to characterize the selectivity of arabinosidases from Lactobacillus brevis. For visualization, antibody-based fluorescent labeling of arabinoxylan coupled with confocal laser-scanning microscopy was established. This allowed for spatial resolution of arabinoxylan within a wheat bran sample and the characterizatioof the effects of several pretreatments. Finally, a rapid screening method based on near and mid-range infrared spectroscopy with multivariate calibration was developed for prediction of nutritional parameters of wheat bran, intended to facilitate income control. These parameters were: water, protein, ash, starch, insoluble as well as soluble dietary fiber and lipids. The established techniques were applied in a concerted effort for the thorough elucidation and assessment of changes in wheat bran and fractions thereof. Additionally, several syntheses of a fluorescent label to directly measure oxidative damage perpetrated on polysaccharides (as e.g. caused by pretreatments) were tested, and are comparatively discussed to serve as a basis for future investigations.