Objective: To evaluate the precision and accuracy of dual‐energy X‐ray absorptiometry (DXA) for the measurement of total‐bone mineral density (TBMD), total‐body bone mineral (TBBM), fat mass (FM), and bone‐free lean tissue mass (LTM) in mice.

Research Methods and Procedures: Twenty‐five male C57BL/6J mice (6 to 11 weeks old; 19 to 29 g) were anesthetized and scanned three times (with repositioning between scans) using a peripheral densitometer (Lunar PIXImus). Gravimetric and chemical extraction techniques (Soxhlet) were used as the criterion method for the determination of body composition; ash content was determined by burning at 600°C for 8 hours.

Results: The mean intraindividual coefficients of variation (CV) for the repeated DXA analyses were: TBMD, 0.84%; TBBM, 1.60%; FM, 2.20%; and LTM, 0.86%. Accuracy was determined by comparing the DXA‐derived data from the first scan with the chemical carcass analysis data. DXA accurately measured bone ash content (p = 0.942), underestimated LTM (0.59 ± 0.05g, p < 0.001), and overestimated FM (2.19 ± 0.06g, p < 0.001). Thus, DXA estimated 100% of bone ash content, 97% of carcass LTM, and 209% of carcass FM. DXA‐derived values were then used to predict chemical values of FM and LTM. Chemically extracted FM was best predicted by DXA FM and DXA LTM [FM = −0.50 + 1.09(DXA FM) − 0.11(DXA LTM), model r² = 0.86, root mean square error (RMSE) = 0.233 g] and chemically determined LTM by DXA LTM [LTM = −0.14 + 1.04(DXA LTM), r² = 0.99, RMSE = 0.238 g].

Discussion: These data show that the precision of DXA for measuring TBMD, TBBM, FM, and LTM in mice ranges from a low of 0.84% to a high of 2.20% (CV). DXA accurately measured bone ash content but overestimated carcass FM and underestimated LTM. However, because of the close relationship between DXA‐derived data and chemical carcass analysis for FM and LTM, prediction equations can be derived to more accurately predict body composition.