Thiophenol is extensively used in the manufacture of industry and agriculture. However, the high concentration of thiophenol will cause damage to living organisms and environment pollution. Herein, a BODIPY-based fluorescent probe has been designed and synthesized for probing thiophenol pollutant in solutions and cells. It shows high selectivity and a turn-on effect. Cell experiments demonstrate that the probe has low cell toxicity, good and rapid cell membrane penetrability, and excellent imaging properties in living cells. The real water samples can be detected directly by fluorescent assay or test strips made from the probe. Therefore, the probe has important value on the detection of thiophenol in the environment and industrial production.