In this work, the sorption and moisture diffusion behaviour of historically relevant glutin-based adhesives (i.e. bone glue, hide glue, fish glue) is characterized. The adhesives sorption isotherms were assessed on thin film samples revealing fundamental differences between the glutin-based adhesives and the synthetic reference adhesive (polyurethane). Furthermore, the water vapour diffusion parallel to the fibre was examined by means of neutron imaging on bonded two-layer samples of Norway spruce wood. In contrast to previous studies using neutron imaging, a new evaluation approach is presented, which allows for nonzero initial moisture conditions and takes into account and compensates for the geometry changes in the sample caused by swelling and shrinkage, thus allowing for a characterization of the diffusion behaviour within the glue line. The diffusion coefficients determined with neutron imaging were interpreted in terms of a theoretical model which takes into account the glue line microstructure. Although the diffusion coefficients were on average larger values for the glutin-based adhesives compared to the reference polyurethane adhesive, the significant variation observed in the sorption measurement is not reflected. This can partially be ascribed to excessive penetration of the adhesives into the wood substrate in fibre direction, which impedes a continuous adhesive layer. Furthermore, deformation and densification of the wood structure was assessed in the vicinity of the adhesive joint. This effect can be ascribed to the surface roughness, which results in very high local stresses leading to buckling and deformation of the tracheids. This situation is similar to that found for adhesive joints in or close to the fibre direction such as finger or butt joints.