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Ag (I)/gelatin microgels with different Ag (I) content have been prepared through self-assembly at various pH values. Silver cations are supposed to be partly trapped in the microgels core after interaction with the carboxyl groups on gelatin chains due to electrostatic attraction. The hydrophobility of gelatin chains enhances when the Ag (I) concentration increases, which results in the contraction of the complex and relatively compactable structure as well as ununiform distribution. The fluorescence experiments show that static quenching plays a main role in the complex formation and the calculated binding sites indicate that pH 7.0—8.0 are optimal for Ag (I)/gelatin self-assembly.