We quantitatively reinvestigated the cells of origin of the spinothalamic tract (STT) of the rat. Injections of Fluoro-Gold that filled the thalamus on one side labeled large numbers of neurons throughout the length of the spinal cord. In 3 cases, we estimated the total number of STT neurons by counting labeled neurons in 18 of the 34 total segments, applying correction factors to these counts, and estimating the numbers of labeled neurons in the 16 remaining unexamined segments. The accuracy of these estimates was tested in two animals in which labeled neurons were counted in all 34 spinal segments. In both cases, the estimated totals of STT neurons differed from the counted totals by less than 5%. In the most effective case, we estimated that more than 9500 STT neurons were labeled. This study indicates that the number of STT neurons in rats is larger than previously reported and suggests that the STT may play an important role in nociception in rats, as it does in primates including humans.