Volumetric assessment and longitudinal changes of brain structures in formalinized Beagle brains

High field MRI represents an advanced technique both for diagnostic and research purposes on animal models such as the Beagle dog. The increasing interest in non-invasive neuroscience, aging, and neuropathological research led to a need of reference values (in terms of volumetric assessment) for the typical brain structures involved and, nowadays, several canine brain MRI atlases have been provided. Since no reports are available regarding the measurements reproducibility and few information are available about formalin fixation effect on brain structures when applied to MRI segmentation, we assessed the segmentation variability of selected structures as a function of the operator (two operators segmented the same data) and their intrinsic variability within a sample of 11 Beagle dogs (9 females and 2 males, 1.6 ± 0.2 years). Then, we analyzed for one further Beagle dog (2 years old) the longitudinal changes in the brain segmentations of these structures corresponding four conditions: in vivo, post mortem (after euthanasia), ex vivo (brain extracted and studied after 1 month in formalin and after 11 months); all the MRI images were collected with a 3 T MRI scanner. Our findings suggest that the segmentation procedure can be considered overall reproducible since only slight statistical differences were detected, apart from the ventricles. Furthermore, in the post mortem/ ex vivo comparison, the majority of the structures showed a higher contrast leading to more reproducible segmentations across operators and a net increase of volume of the studied structures; this could be justified by the intrinsic relaxation time changes observed as a consequence of formalin fixation, that led to an improvement of brain structures visualization and then segmentation. To conclude, MRI based segmentation seems to be a useful and accurate tool that allows longitudinal studies, especially when applied to formalin fixed brains. ### Competing Interest Statement The authors have declared no competing interest.