1：Experimental Design and Method Selection

The study involved the synthesis of red-fluorescent amphiphilic block copolymers (RPO-1–3) with different hydrophobic blocks (PCL and PDL) and hydrophilic blocks (POEGMA). The self-assembly of these polymers in aqueous solutions was characterized using transmission electron microscopy (TEM). The drug-loading capacity and release kinetics were studied using doxorubicin (DOX) as a model anticancer drug.

2：Sample Selection and Data Sources

The samples included HeLa cells for cytotoxicity studies and the polymers RPO-1, RPO-2, and RPO-3 for drug delivery and imaging studies.

3：List of Experimental Equipment and Materials

Transmission electron microscope (TEM), UV–vis absorption spectroscopy, fluorescence spectroscopy, dynamic light scattering (DLS) instrument, confocal laser scanning microscopy (CLSM).

4：Experimental Procedures and Operational Workflow

The synthesis of polymers involved ring-opening polymerization and atom transfer radical polymerization (ATRP). The self-assembly of polymers was induced by slow addition of water into polymer solutions. Drug encapsulation and release studies were conducted using dialysis methods. Cellular uptake and cytotoxicity were assessed using CLSM and PrestoBlue assay.

5：Data Analysis Methods

Data analysis included characterization of polymer structures using NMR and GPC, optical properties using UV–vis and fluorescence spectroscopy, and drug release profiles using fluorescence intensity measurements.