We investigate the influence of various optical fiber fabrication processes on the fluorescence decay of RE ions commonly used in fiber lasers and amplifiers, i.e. Yb3+, Tm3+ and Ho3+. Optical fiber preforms were prepared using the MCVD method combined with Al2O3 nanoparticle doping and subjected to subsequent heat treatment processes such as preform elongation and fiber drawing. The fluorescence decay of RE ions was measured in multiple stages of optical fiber preparation: in an original preform, in an elongated preform (cane), in a standard fiber, and in an overcladded fiber. It was found that heat treatment processing of the preforms generally leads to a faster fluorescence decay, which can be explained by the diffusion of dopants and clustering of RE ions. The fiber drawing exhibited a greater effect compared to preform elongation, which was ascribed to a faster cooling rate of the process. In general, the heat treatment of RE-doped silica glass preforms leads to the decline of fluorescence decay.