Synthesis and Characterization of Amphiphilic AIE Polymersomes

Amphiphilic block-co-polymers PEG-P(AIE) were synthesized according to the previously reported methodology [22 (link)]. To prepare the AIE polymersomes, PEG₄₄-P(AIE)₁₄ was dissolved in THF (2 mg/mL) in a 4 mL glass vial with a magnetic stirring bar and sealed with a rubber septum. After stirring for 10 min, 0.5 mL of ultrapure Milli-Q water was added to the polymer solution through a syringe pump (Chemyx, Inc., Fusion 100, KR Analytical Limited, Stafford, TX, USA) with a speed of 0.25 mL/h. Then, the resulting cloudy solution was transferred into a pre-hydrated dialysis bag (Spectra/Pro^®, MWCO 12,000–14,000, 2 mL/cm, Rancho Dominguez, CA, USA) for dialysis against 0 mM NaCl or 100 mM NaCl solution at 4 °C for at least 24 h. Dynamic light scattering (DLS, Nano ZSP, Malvern Instruments, Malvern, UK), scanning electron microscopy (SEM, FEI Quanta 200 3D FEG, Thermo Fisher Scientific, Waltham, MA, USA), and cryogenic transmission electron microscopy (cryo-TEM, TU/e CryoTitan, Thermo Fisher Scientific, Waltham, MA, USA) were used for size and morphological characterization. For characterization of the fluorescence, a Spark^® 10M microplate reader (TECAN, Männedorf, Switzerland), and two-photon/confocal laser scanning microscopy (TP-CLSM, Leica TCS SP8X, Wetzlar, Germany) were applied.