To correlate thermal dose from focused ultrasound (FUS) with gene expression and tissue injury, a temperature plateau strategy was employed. Plasmids encoding luciferase gene under the control of hsp70B promoter were transfected into the right gastrocnemius muscle in a rat via electroporation. One day after transfection, hind limbs were treated with 3.3-MHz focused ultrasound, using one of four different temperature plateaus with spatial-peak time-average focal temperatures (T(SPTA)) of 46 degrees C, 48 degrees C, 51 degrees C and 62 degrees C. The treatment duration at the plateau temperature was varied from 0 to 30s. Gene expression was analyzed in vivo one day following FUS treatment, and H&E staining was employed to assess tissue injury. Gene activation and tissue damage correlated closely with thermal dose. The highest level of gene activation was induced by FUS at T(SPTA)=51 degrees C for 20s, which was found to be statistically equivalent to that produced by water-bath hyperthermia.