The application of different sensing principles in microfluidic devices opens up further possibilities for the development of point-of-care testing (POCT). Herein, the photothermal sensing principle is introduced in microfluidic paper-based analytical devices (μPADs) to develop a photothermal microfluidic sensing platform using near-infrared (NIR) laser-driven multiplexed dual-mode visual quantitative readout. Prussian blue (PB) as the analyte-associated photothermal agent was in situ synthesized in thermoresponsive poly(N-isopropylacrylamide) hydrogels to serve as the on-chip photothermal sensing element. The NIR laser-driven photothermal effect of PB triggered not only on-chip dose-dependent heat generation but also phase transition-induced dye release from the hydrogels, simultaneously enabling both thermal image- and distance-based dual-mode visual quantitative readout of the analyte concentration in a multiplexed manner. Both the on-chip temperature elevation value of the hydrogels and the traveling distance of released dye solutions were proportional to the concentration of PB. With the detection of silver ions in environmental water as a proof-of-concept study, the photothermal μPAD can detect silver ions at a concentration as low as 0.25 μM with high selectivity and satisfactory accuracy. The photothermal microfluidic sensing platform holds great potential for POCT with promising integratability and broad applicability, owing to the combination of synergistic advantages of the photothermal sensing principle, μPADs, and photothermally responsive hydrogels.