Poly(N‐isopropylacrylamide) (PNIPAM) hydrogel microspheres were prepared by precipitation polymerization above the polymer's lower critical solution temperature (LCST) in water. The hydrodynamic size of PNIPAM microspheres exhibited a thermo‐sensitive change around the LCST. In order to obtain information about the interface of the hydrogels, contact angle measurement, protein adsorption, and fluorescence analysis were performed. Contact angle measurement and protein adsorption showed a gradual change around the LCST similar to the change of the hydrodynamic size. However, fluorescence study exhibited a sharp change at the LCST. In addition, a quenching experiment was employed to elucidate the location of the fluorescent probes. In the PNIPAM latex, the probes were located where they were less quenchable as compared to the PNIPAM solution. It seems probable that the probes can penetrate into the crosslinked PNIPAM hydrogel and might reflect the inner environment of the hydrogel. The effect of salt on thermo‐sensitive behaviour was also detected by fluorescence analysis. On the other hand, the hydrogel microsphere prepared from poly(acryloyl pyrrolidine) (PAPr) showed a gradual change in fluorescence with temperature.
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