Tomographic reconstruction of rat glomerular capillary glycocalyx using LaDy GAGa staining on a 300 nm thick section. A-D: raw transmission electron microscopy at -60o,-30o,30o and 60o respectively (Additional file 14: Movie S7A). E-H: reconstructed images of glycocalyx in cross-section 30 nm apart (Additional file 15: Movie S7B). There is the appearance of a gap between the glycocalyx (Gly) and the endothelial cell (En). Small holes can be seen throughout Gly, these are gaps between the fibres. Boxes are examples of areas used for spacing analysis. I-L: are the reconstruction viewed perpendicular to the membrane. I: is at the top of the glycocalyx, J: is 70 nm from the top, K: is 140 nm from the top and L: is within the endothelial layer (fenestrations can clearly be seen). M-O: Are stills from the 3D projection (Additional file 16: Movie S7C). A-L: scale bar is 300 nm. M-O: scale bar is 150 nm. Labels: Cap = capillary lumen, Gly = glycocalyx, Fen = Fenestration, GBM = glomerular basement membrane, SPS = subpodocyte space, Slit = Slit Diaphram (the diaphragm is discernable), Pod = Podocyte (foot process), En = endothelial cell.