OBJECTIVE: To assess the accuracy and reproducibility of dual-energy absorptiometry (DXA; PIXImus(™)) and time domain nuclear magnetic resonance (TD-NMR; Bruker Optics) for the measurement of body composition of lean and obese mice. SUBJECTS AND MEASUREMENTS: Thirty lean and obese mice (body weight range 19-67 g) were studied. Coefficients of variation for repeated (x 4) DXA and NMR scans of mice were calculated to assess reproducibility. Accuracy was assessed by comparing DXA and NMR results of ten mice to chemical carcass analyses. Accuracy of the respective techniques was also assessed by comparing DXA and NMR results obtained with ground meat samples to chemical analyses. Repeated scans of 10-25 gram samples were performed to test the sensitivity of the DXA and NMR methods to variation in sample mass. RESULTS: In mice, DXA and NMR reproducibility measures were similar for fat tissue mass (FTM) (DXA coefficient of variation [CV]=2.3%; and NMR CV=2.8%) (P=0.47), while reproducibility of lean tissue mass (LTM) estimates were better for DXA (1.0%) than NMR (2.2%) (<P 0.05). Regarding accuracy, in mice, DXA overestimated (vs chemical composition) LTM (+1.7 ± 1.3 g [SD], ~ 8%, P <0.001) as well as FTM (+2.0 ± 1.2 g, ~ 46%, P <0.001). NMR estimated LTM and FTM virtually identical to chemical composition analysis (LTM: -0.05 ± 0.5 g, ~0.2%, P =0.79) (FTM: +0.02 ± 0.7 g, ~15%, P =0.93). DXA and NMR-determined LTM and FTM measurements were highly correlated with the corresponding chemical analyses (r(2)=0.92 and r(2)=0.99 for DXA LTM and FTM, respectively; r(2)=0.99 and r(2)=0.99 for NMR LTM and FTM, respectively.) Sample mass did not affect accuracy in assessing chemical composition of small ground meat samples by either DXA or NMR. CONCLUSION: DXA and NMR provide comparable levels of reproducibility in measurements of body composition lean and obese mice. While DXA and NMR measures are highly correlated with chemical analysis measures, DXA consistently overestimates LTM and FTM (by ~8% and ~46%, respectively), while NMR only slightly underestimates LTM (by ~0.2%) and overestimates FTM (~15%.) The NMR method also has practical advantages compared to DXA, such as speed of measurement and the ability to scan unanesthetized animals.