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Conjugated polymer nanoparticles, produced by in situ colloidal Knoevenagel polymerization, show advantageous properties (bright emission, colloidal/chemical stability and mesoscopic size range) that allow the successful in vivo application to real-time sentinel lymph node mapping in a mouse model.

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Hydrogen peroxide (H(2)O(2)) is an endogenous molecule that plays diverse physiological and pathological roles in living systems. Here we report multimolecule integrated nanoprobes with the enhanced chemiluminescence (CL) response to H(2)O(2) that is produced in cells and in vivo. This approach is based on the nanoscopic coaggregation of a dye exhibiting aggregation-enhanced fluorescence (AEF) with a H(2)O(2)-responsive peroxalate that can convert chemical reaction energy into electronic excitation. The coaggregated CL nanoparticles (FPOA NPs) with an average size of ~20 nm were formulated by aqueous self-assembly of a ternary mixture of a surfactant (Pluronic F-127) and concentrated hydrophobic dye/peroxalte payloads. Spectroscopic studies manifest that FPOA NPs as a reagent-concentrated nanoreactor possess the signal enhancement effect by AEF, as well as the optimized efficiencies for H(2)O(2) peroxalate reaction and subsequent intraparticle energy transfer to the dye aggregates, to yield greatly enhanced CL generation with a prolonged lifetime. It is shown that the enhanced CL signal thereby is capable of detecting intracellular H(2)O(2) overproduced during immune response. We also demonstrate that the densely integrated nature of FPOA NPs facilitates further intraparticle CL energy transfer to a low-energy dopant to red shift the spectrum toward the biologically more transparent optical window, which enables the high-sensitivity in vivo visualization of H(2)O(2) associated with early stage inflammation.