Dual-axis electron tomography: a new approach for investigating the spatial organization of wood cellulose microfibrils

We have employed dual-axis electron tomography to investigate the 3D organization of cellulose microfibrils in plastic resin-embedded, delignified cell walls of radiata pine early wood. The ∼ 1 nm thick tomographic slices produced in this study provided for a resolution of ∼ 2 nm in the cross-section of the slices throughout the 150 nm thick plastic sections. This resolution is sufficient to resolve individual cellulose microfibrils and to map the 3D organization of the cellulose microfibrils within the S2 layer of the secondary cell walls. The individual cellulose microfibrils measure ∼ 3.2 nm in diameter, and appear to consist of a ∼ 2.2 nm unstained core and a ∼ 0.5 nm thick surface layer that is lightly stained. Both individual and clustered cellulose microfibrils are seen surrounded by more heavily stained and irregularly shaped residual lignin and hemicellulose. The tightness of packing of the cellulose microfibrils in the cluster varies along the thickness of the section. These findings demonstrate that dual-axis electron tomography is a technique that can provide new insights into the 3D organization of cellulose microfibrils in plant cell walls.