Four metal speciation and fractionation techniques – DGT (diffusive gradients in thin films), 1-kDa ultrafiltration, 0.22-µm membrane filtration and aquatic moss – were simultaneously applied to a small, contaminated freshwater stream in northern Sweden to investigate differences and similarities between the methods regarding trace metal speciation and their dependence on geochemical water properties. The investigated metals comprise Al, Cd, Co, Cu, Fe, Mn, Ni, and Zn. The normal DGT devices with Chelex cation exchanger were used. Shoots from the aquatic moss Fontinalis antipyretica L ex Hedw. were collected in a non-polluted brook and transplanted to the sampling site for exposure. It was evident that 0.22-µm membrane filtration, 1-kDa ultrafiltration and DGT generally measured different metal fractions where <1-kDa ultrafiltered concentrations were lower than DGT labile concentrations which in turn were lower than <0.22-µm concentrations. The differences between DGT and <1-kDa concentrations indicate the occurrence of labile colloids discriminated by ultrafiltration. Strong correlations between DGT and <1 kDa concentrations were found for Al, Cu, Cd, Co and Zn. Despite a rigorous sample cleaning, retention of particulate matter on the moss samples was revealed by a significant correlation between metal concentrations in moss and particulate Fe. Generally, elevated trace metal concentrations were found in moss exposed at the sampling site compared to reference moss from the non-polluted brook. No significant correlations were found between DGT-labile concentrations and moss concentrations.