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Double diffusion encoding (DDE) magnetic resonance measurements of the watersignal offers a unique ability to separate the effect of microscopicanisotropic diffusion in structural units of tissue from the overallmacroscopic orientational distribution of cells. However, the specificity indetected microscopic anisotropy is limited as the signal is averaged overdifferent cell types and across tissue compartments. Performing side-by-sidemetabolite DDE spectroscopy (DDES) and water DDES in which a wide range ofb-values is used to gradually eliminate the extracellular contribution providescomplementary measures from which intracellular and extracellular microscopicfractional anisotropies ($\mu$FA) and diffusivities can be estimated.Metabolites are largely confined to the intracellular space and thereforeprovide a benchmark for intracellular diffusivity of specific cell types. Here,we aimed to estimate tissue- and compartment-specific human brainmicrostructure by combining water and metabolites DDES experiments. Weperformed DDES in human subjects in two brain regions that contain widelydifferent amounts of white matter (WM) and gray matter (GM): parietal whitematter (PWM) and occipital gray matter (OGM) on a 7 T MRI scanner. Results ofthe metabolite DDES experiments in both PWM and OGM suggest a highlyanisotropic intracellular space within neurons and glia, with the possibleexception of gray matter glia. Tortuosity values in the cytoplasm for water andtNAA, obtained with correlation analysis of microscopic parallel diffusivitywith respect to GM/WM tissue fraction in the volume of interest, are remarkablysimilar for both molecules, while exhibiting a clear difference between grayand white matter, suggesting a more crowded cytoplasm and more complexcytomorphology of neuronal cell bodies and dendrites in GM than those found inlong-range axons in WM.

Compartmental diffusion and microstructural properties of human brain  gray and white matter studied with double diffusion encoding magnetic  resonance spectroscopy of metabolites and water