Quantitative in vivo micro-computed tomography for assessment of age-dependent changes in murine whole-body composition

Micro-computed tomography (micro-CT) is used routinely to quantify skeletal tissue mass in small animal models. Our goal was to evaluate repeated in vivo micro-CT imaging for monitoring whole-body composition in studies of growth and aging in mice. Male mice from 2 to 52 weeks of age were anesthetized and imaged using an eXplore Locus Ultra and/or eXplore speCZT scanner. Images were reconstructed into 3D volumes, signal-intensity thresholds were used to classify each voxel as adipose, lean or skeletal tissue, and tissue masses were calculated from known density values. Images revealed specific changes in tissue distribution with growth and aging. Quantification showed biphasic increases in total CT-derived body mass, lean and skeletal tissue masses, consisting of rapid increases to 8 weeks of age, followed by slow linear increases to 52 weeks. In contrast, bone mineral density increased rapidly to a stable plateau at ~ 14 weeks of age. On the other hand, adipose tissue mass increased continuously with age. A micro-CT-derived total mass was calculated for each mouse and compared with gravimetrically measured mass, which differed on average by < 3%. Parameters were highly reproducible for mice of the same age, but variability increased slightly with age. There was also good agreement in parameters for the same group of mice scanned on the eXplore Locus Ultra and eXplore speCZT systems. This study provides reference values for normative comparisons; as well, it demonstrates the usefulness of in vivo single-energy micro-CT scans to quantify whole-body composition in high-throughput studies of growth and aging in mice.