Imaging and reconstruction of the cytoarchitecture of axonal fibres: enabling biomedical engineering studies involving brain microstructure

Abstract There is an increased need and focus to understand how local brain microstructure affects the transport of drug molecules directly administered to the brain tissue, for example in convection-enhanced delivery procedures. This study reports the first systematic attempt to characterize the cytoarchitecture of commissural, long association and projection fibers, namely: the corpus callosum, the fornix and the corona radiata. Ovine samples from three different subjects were stained with osmium tetroxide (to enhance contrast from cell organelles and the fibers), embedded in resin and then imaged using scanning electron microscope combined with focused ion beam milling to generate 3D volume reconstructions of the tissue at subcellular spatial resolution. Particular focus has been given to the characteristic cytological feature of the white matter: the axons and their alignment in the tissue. Via 2D images a homogeneous myelination has been estimated via detection of ~40% content of lipids in all the different fiber tracts. Additionally, for each tract, a 3D reconstruction of relatively large volumes (15μm x 15μm x 15μm – including a significant number of axons) has been performed. Namely, outer axonal ellipticity, outer axonal cross-sectional area and their relative perimeter have been measured. The study of well-resolved microstructural features provides useful insight into the fibrous organization of the tissue, whose micromechanical behaviour is that of a composite material presenting elliptical tortuous tubular fibers embedded in the extra-cellular matrix. Drug flow can be captured through microstructurally-based models, leading to a workflow to enable physically-accurate simulations of drug delivery to the targeted tissue.