The sol-gel transition of aqueous barley beta-glucan solutions which undergo gelation with ageing has been studied by conventional bulk rheology, phase contrast microscopy and particle tracking microrheology. Characterisation of the primary structure of the beta-glucan isolate was carried out by enzymic methods and HPLC. The Brownian diffusion of fluorescent microspheres (0.75 mu m diameter, carboxylate-coated particles) was used to probe the spatial mechanical properties of the gelling systems at the scale of microns; the potential use of passive particle tracking to study biopolymer gelling systems that present spatial heterogeneities is thus explored. For the beta-glucan gels cured at 25 degrees C both microrheology and bulk rheology revealed that with increasing the polysaccharide concentration the gelation time decreased, while the gelation rate and gel strength of the barley beta-glucan gels increased. The particle tracking method had higher sensitivity and could map molecular ordering and structural heterogeneities in the evolving polysaccharide network at a micro-level. That is, different size pores were generated upon ageing with regions of depleted or less amount of beta-glucan molecules. Furthermore, this method could detect changes in the fine structure of the system before such events can be registered by bulk rheological measurements; i.e. microheterogeneity and aggregation of beta-glucan chains were revealed by particle tracking at earlier temporal stages of the experiment. (C) 2012 Elsevier Inc. All rights reserved.