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Background:  Brainstem-mediated functions are impaired in neurodegenerative diseases and aging. Atrophy can be visualized by MRI. This study investigates extrinsic sources of brainstem volume variability, intrinsic sources of anatomical variability, and the influence of age and sex on the brainstem volumes in healthy subjects. We aimed to develop efficient normalization strategies to reduce the effects of intrinsic anatomic variability on brainstem volumetry.   Methods:  Brainstem segmentation was performed from MPRAGE data using our deep-learning-based brainstem segmentation algorithm MD-GRU. The extrinsic variability of brainstem volume assessments across scanners and protocols was investigated in two groups comprising 11 (median age 33.3 years, 7 women) and 22 healthy subjects (median age 27.6 years, 50% women) scanned twice and compared using Dice scores. Intrinsic anatomical inter-individual variability and age and sex effects on brainstem volumes were assessed in segmentations of 110 healthy subjects (median age 30.9 years, range 18–72 years, 53.6% women) acquired on 1.5T (45%) and 3T (55%) scanners. The association between brainstem volumes and predefined anatomical covariates was studied using Pearson correlations. Anatomical variables with associations of | r | > 0.30 as well as the variables age and sex were used to construct normalization models using backward selection. The effect of the resulting normalization models was assessed by % relative standard deviation reduction and by comparing the inter-individual variability of the normalized brainstem volumes to the non-normalized values using paired t- tests with Bonferroni correction.   Results:  The extrinsic variability of brainstem volumetry across different field strengths and imaging protocols was low (Dice scores > 0.94). Mean inter-individual variability/SD of total brainstem volumes was 9.8%/7.36. A normalization based on either total intracranial volume (TICV), TICV and age, or v-scale significantly reduced the inter-individual variability of total brainstem volumes compared to non-normalized volumes and similarly reduced the relative standard deviation by about 35%.   Conclusion:  The extrinsic variability of the novel brainstem segmentation method MD-GRU across different scanners and imaging protocols is very low. Anatomic inter-individual variability of brainstem volumes is substantial. This study presents efficient normalization models for variability reduction in brainstem volumetry in healthy subjects.

Improving Accuracy of Brainstem MRI Volumetry: Effects of Age and Sex, and Normalization Strategies