Luminescence-enhanced conjugated polymer dots through thermal treatment for cell imagingElectronic supplementary information (ESI) available. See DOI: https://doi.org/10.1039/d2bm00516f

Conjugated polymer dots (Pdots) are often used as excellent fluorescent probes in the biomedical field. In the process of preparing Pdots, the rapid change of the solvent polarity will result in a messy and defective stacking of the polymer chains in the particle, and these stacking defects of the polymer chains may weaken its luminescence properties. Here, we try to optimize the stacking of the conjugated polymer chains by the thermal annealing treatment. After the low temperature thermal treatment, the fluorescence intensity of Pdots can be enhanced by about 11%–29%, and Pdots maintain their original stability and biosafety. We used transmission electron microscopy (TEM) and single particle fluorescence imaging to reveal the possible mechanism of the chain stacking optimization process, that is, the thermal annealing process of Pdots is the competition between internal chain rearrangement in the particle and particle aggregation. The luminescence-enhanced Pdots exhibit good cellular imaging performance. These results prove that it is feasible to extend the thermal annealing treatment from planar polymer devices to polymer nanoparticles. It provides the possibility to realize stable and complex biological imaging applications using Pdots with a simple molecular structure, and a mature improvement scheme for the mass preparation of Pdots.