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Purpose To introduce in vivo multifrequency single‐shot magnetic resonance elastography for full‐FOV stiffness mapping of the mouse brain and to compare in vivo stiffness of neural tissues with different white‐to‐gray matter ratios. Methods Viscous phantoms and 10 C57BL‐6 mice were investigated by 7T small‐animal MRI using a single‐shot spin‐echo planar imaging magnetic resonance elastography sequence with motion‐encoding gradients positioned before the refocusing pulse. Wave images were acquired over 10 minutes for 6 mechanical vibration frequencies between 900 and 1400 Hz. Stiffness maps of shear wave speed (SWS) were computed using tomoelastography data processing and compared with algebraic Helmholtz inversion (AHI) for signal‐to‐noise ratio (SNR) analysis. Different brain regions were analyzed including cerebral cortex, corpus callosum, hippocampus, and diencephalon. Results In phantoms, algebraic Helmholtz inversion–based SWS was systematically biased by noise and discretization, whereas tomoelastography‐derived SWS was consistent over the full SNR range analyzed. Mean in vivo SWS of the whole brain was 3.76 ± 0.33 m/s with significant regional variation (hippocampus = 4.91 ± 0.49 m/s, diencephalon = 4.78 ± 0.78 m/s, cerebral cortex = 3.53 ± 0.29 m/s, and corpus callosum = 2.89 ± 0.17 m/s). Conclusion Tomoelastography retrieves mouse brain stiffness within shorter scan times and with greater detail resolution than classical algebraic Helmholtz inversion–based magnetic resonance elastography. The range of SWS values obtained here indicates that mouse white matter is softer than gray matter at the frequencies investigated.

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