The objective of this study was to determine the effect of fixation, soft tissues, and scan projection on bone mineral measurements with dual energy X-ray absorptiometry (DXA). In seven fresh cadavers, DXA scans were obtained within 48 hours of death and after 10 months of fixation with 5% formalin/95% ethanol. The measurements showed a high linear relationship (r2 > 0.97; SEE% 0.90; SEE% < 9%), but led to more substantial deviations in regional femoral analyses and in the spine (r2 = 0.78-0.90; SEE% = 8-22%). Comparing lateral with anterior-posterior (AP) spinal scans, the vertebral bodies were found to occupy 40.2 +/- 7.2% of the BMC, and 62.0 +/- 11.2% of the BMD, the ranges being 26-58%, and 38-91%, respectively. There were large deviations from linearity between in situ AP and ex situ lateral spinal scans with r2 values of 0.63 and 0.73 for BMD and BMC (SEE% = 52% and 27% relative to the vertebral body), respectively.