We report here simple synthetic method for preparing polymer dots (Pdots) via hydrothermal treatment of organic dye (neutral red), urea and trisodium citrate. The prepared Pdots with enhanced quantum yield (quantum yield: 30.2%) was used as a selective and sensitive probe for fluorescent sensing of dopamine (DA) with high selectivity and sensitivity. The as-synthesized Pdots exhibited strong fluorescence intensity at 435 nm, which DA can trigger remarkable fluorescence quenching of such luminescent Pdots on the basis of inner filter effect (IFE) and static quenching effect (SQE). A wide linearity range (0.001 mMe900 mM) for DA detection was obtained with lower DL (3 S/N) of 0.28 nM, and no interference from other molecules such as ascorbic acid, urine acid, glutathione, glucose, epinephrine, arginine, cysteine, proline, creatinine, serine; alanine, L-therionine, Hg2þ, Mg2þ, Kþ, Ca2þ and Naþ. The designed sensor was successfully applied in the imaging of DA in single living PC12 cells using droplet microfluidic approach, indicating its acceptable practicability of the proposed assay for DA detection with ultrahigh sensitivity in biological samples.