In vivo rat brain mapping of multiple gray matter water populations using nonparametric D(ω)-R₁-R₂ distributions MRI

Massively multidimensional diffusion magnetic resonance imaging combines tensor-valued encoding, oscillating gradients, and diffusion-relaxation correlation to provide multicomponent subvoxel parameters depicting some tissue microstructural features. This method was successfully implemented ex vivo in microimaging systems and clinical conditions with tensor-valued gradient waveform of variable duration giving access to a narrow diffusion frequency (ω) range. We demonstrate here its preclinical in vivo implementation with a protocol of 389 contrast images probing a wide diffusion frequency range of 18 to 92 Hz at b-values up to 2.1 ms/μm² enabled by the use of modulated gradient waveforms and combined with multislice high-resolution and low-distortion echo planar imaging acquisition with segmented and full reversed phase-encode acquisition. This framework allows the identification of diffusion ω-dependence in the rat cerebellum and olfactory bulb gray matter (GM), and the parameter distributions are shown to resolve two water pools in the cerebellum GM with different diffusion coefficients, shapes, ω-dependence, relaxation rates, and spatial repartition whose attribution to specific microstructure could modify the current understanding of the origin of restriction in GM.