Hyperthermia has been used as a cancer treatment in which tumors are elevated to cytotoxic temperatures to aid in their control. A noninvasive method for volumetrically determining temperature distribution during treatment would greatly enhance the ability to uniformly heat tumors at therapeutic levels. Ultrasound is an attractive modality for this purpose. We investigated changes in backscattered energy (CBE) from pulsed ultrasound with temperature. Our predicted changes in backscattered energy were matched by in vitro measurements in samples of bovine liver, turkey breast, and pork rib muscle. We studied CBE in tissue regions with multiple scatterers, of isolated individual scatterers, and in collections of individual scatterers. The latter appears to have the most potential. We measured the CBE with a focused circular transducer with a center frequency of 7.5 MHz. The standard deviation of the CBE of 75-125 scattering regions from 0.3 to 0.5 cm3 volumes increased nearly monotonically from 37 degrees C to 50 degrees C in each tissue type. Although the slopes were different, the curve for each type of tissue was well matched by a second-degree polynomial, with a correlation coefficient of 0.99 in each case. Thus the use of the CBE of ultrasound for temperature estimation may have clinical promise with a convenient, low cost modality. Because our approach exploits the inhomogeneities present in tissue, we believe that if it is successful in vitro, it holds promise for in vivo application.