Tuning the Internal Compartmentation of Single‐Chain Nanoparticles as Fluorescent Contrast Agents

Controlling the internal structures of single‐chain nanoparticles (SCNPs) is an important factor for their targeted chemical design and synthesis, especially in view of nanosized compartments presenting different local environments as a main feature to control functionality. We here design SCNPs bearing near‐infrared fluorescent dyes embedded in hydrophobic compartments for use as contrast agents in pump–probe photoacoustic (PA) imaging, displaying improved properties by the location of the dye in the hydrophobic particle core. Compartment formation is controlled via single‐chain collapse and subsequent crosslinking of an amphiphilic polymer using external crosslinkers in reaction media of adjustable polarity. Different SCNPs with hydrodynamic diameters of 6‐12 nm bearing adjustable label densities are synthesized. It is found that the specific conditions for single‐chain collapse have a major impact on the formation of the desired core–shell structure, in turn adjusting the internal nanocompartments together with the formation of excitonic dye couples, which in turn increase their fluorescence lifetime and PA signal generation. SCNPs with the dye molecules accumulate at the core also show a nonlinear PA response as a function of pulse energy—a property that can be exploited as a contrast mechanism in molecular PA tomography. An amphiphilic polymer is collapsed into single‐chain nanoparticles in either water or tetrahydrofuran using a crosslinking NIR‐fluorescent dye with a non‐functional crosslinker. The formation of the desired nano‐compartmented structures is studied together with the usefulness of these functionalized nanoparticles as contrast agent for pump–probe photoacoustic imaging.