Although there are several techniques currently available to assess skeletal integrity in live birds, few offer important features for application in a commercial setting, offering ease of use and moderate cost. Quantitative ultrasonography (QUS) is an established technique for diagnosis of osteoporosis in humans and horses that has potential application for layers. An OmniSense 700S quantitative ultrasonometer was evaluated for use with Single Comb White Leghorn hens. Humeral QUS values (m/s) were measured in a series of experiments using a total of 144 Shaver White hens. Significant correlations (P < 0.01) were observed among sequential QUS measurements taken on the same bird at 54, 60, and 66 wk of age. At the completion of the studies (66 wk of age), the left and right humeri were excised, cleaned, and rescanned. Postmortem QUS data from left and right humeri were related (R2 = 0.72, P < 0.0001), although future studies may need to consider both sides of the skeleton to account for asymmetry conditions. Ultrasound data collected from live hens at 66 wk of age correlated well with postmortem QUS data (R2 = 0.80, P < 0.0001). Quantitative ultrasonography did not correlate with humeral bone-breaking force measured postmortem. Bones from live hens, surrounded by tissue thicker than 4 mm, could not be read by the QUS probe.