Hydrogen sulfide (H2S) is an important signaling molecule in various biological processes; however, its real-time monitoring in living cells is hampered by long detection time for current fluorescent probes. To overcome this challenge, we designed a phase-transfer catalyst (PTC) approach to accelerate the reaction between the probe and the analyte by conjugating common fluorescent probes-mostly hydrophobic small molecules-with an amphiphilic PEG-PPG-PEG polymer, enabling the controllable assembly of H2S nanoprobes in an aqueous solution. The PEG block helps to establish a PTC microenvironment that endows the assembled nanoprobes with a significantly reduced detection time (3-10 min; versus 20-60 min for small-molecule probes). Based on this approach, we synthesised two nanoprobes of different wavelengths, DS-Blue-nano and DN-Green-nano, which can sensitively detect H2S in living macrophage cells with bright fluorescence starting at as early as 7 min and reaching stability at 15 min. These data suggest PTC-based nanoprobes as a new and generic approach for constructing sensitive fluorescent probes for the real-time imaging of H2S, and perhaps other molecules in future, under biological conditions.