Towards a representative reference for MRI-based human axon radius assessment using light microscopy

Non-invasive assessment of axon radii via MRI bears great potential for clinical and neuroscience research as it is a main determinant of the neuronal conduction velocity. However, there is a lack of representative histological reference data at the scale of the cross-section of MRI voxels for validating the MRI-visible, effective radius (r_eff). Because the current gold standard stems from neuroanatomical studies designed to estimate the bulk-determined arithmetic mean radius (r_arith) on small ensembles of axons, it is unsuited to estimate the tail-weighted r_eff. We propose CNN-based segmentation on high-resolution, large-scale light microscopy (lsLM) data to generate a representative reference for r_eff. In a human corpus callosum, we assessed estimation accuracy and bias of r_arith and r_eff. Furthermore, we investigated whether mapping anatomy-related variation of r_arith and r_eff is confounded by low-frequency variation of the image intensity, e.g., due to staining heterogeneity. Finally, we analyzed the error due to outstandingly large axons in r_eff. Compared to r_arith, r_eff was estimated with higher accuracy (maximum normalized-root-mean-square-error of r_eff: 8.5 %; r_arith: 19.5 %) and lower bias (maximum absolute normalized-mean-bias-error of r_eff: 4.8 %; r_arith: 13.4 %). While r_arith was confounded by variation of the image intensity, variation of r_eff seemed anatomy-related. The largest axons contributed between 0.8 % and 2.9 % to r_eff. In conclusion, the proposed method is a step towards representatively estimating r_eff at MRI voxel resolution. Further investigations are required to assess generalization to other brains and brain areas with different axon radii distributions.