Electrospun polymer fibers have attracted great attention in recent years because of their unique properties and applications in various areas such as tissue engineering and drug delivery. It remains a great challenge, however, to understand the morphology and property changes of electrospun polymer fibers induced by post-treatment, especially thermal annealing. In this work, we investigate the morphology evolution of electrospun polystyrene (PS) fibers thermally annealed on poly(vinyl alcohol) (PVA) films. Driven by the surface and interfacial tensions, the PS fibers transform to a series of regularly aligned anisotropic particles embedded in the PVA films via a Rayleigh instability-type transformation. From the in-situ observed optical microscopy (OM) images, higher growth rates of undulated amplitude can be obtained for thinner PVA films, revealing the effect of the underlying glass substrates. (C) 2017 Elsevier Ltd. All rights reserved.