Thallium-doped cesium iodide (CsI(Tl)) screens are widely used in X-ray imaging devices because of the columnar structure of the CsI(Tl) layer, but few reports focus on the optical role of the substrate in the screen system. In this paper, four substrates including fused silica (SiO2), silver-film coated SiO2, graphite (C) and fiber optic plate (FOP) are used to fabricate CsI(Tl) screens by thermal evaporation. Their imaging performance is evaluated by relative light output (RLO), modulation transfer function (MTF), normalized noise power spectrum (NNPS) and noise equivalent quanta (NEQ). The results reveal that although CsI(Tl) film on graphite plate yields images with the lowest light output, it presents relatively higher spatial resolution and better signal-to-noise characteristics. However, films on SiO2 plate obtain low MTF but high NNPS curves, whether they are coated with silver film or not. Furthermore, scintillation screens on FOP have bright images with low NNPS and high NEQ, but have the lowest MTF. By controlling the substrate optical features, CsI(Tl) films can be tailored to suit a given application.