Thiophenol is known as a class of highly toxic pollutant that can damage to the environment and biological health. It is great significant to detect thiophenol rapidly and sensitively in living cells and environment samples. Hence, we designed and developed two reliable and efficient fluorescent probes that can monitor thiophenol rapidly. The two probes had superior properties in sensitivity and selectivity in the solvent system of pH = 7.4, which had high reactivity to thiophenol, and basically had no response to other sulfur compounds. These probes had a superior range of linearity and low limit of detection for thiophenols. The limits of detection for probe A and probe B (calculated by 3σ/k) are 63.9 nM and 24.7 nM, respectively. These probes have been applied to detect thiophenol in environmental water with good recovery. Moreover, probe B has been used to visualize thiophenol in living cells. Therefore, the fluorescent probes can provide an effective way to monitor thiophenols in environmental water and in cell imaging.