On the basis of the mechanism of Hg(2+)-promoted hydrolysis, a new fluorescent chemodosimeter (Rho-Hg1) is reported for single-selective and parts per billion level-sensitive detection of Hg(2+) in natural waters. Moreover, the fluorescence response of Rho-Hg1 to Hg(2+) has little interference from sulfur compounds such as cysteine and glutathione and could be used in the Hg(2+) imaging in living cells.