The aim of our work was to provide a robust method for evaluating imaging performance of positron emission tomography (PET) systems and particularly to estimate the modulation transfer function (MTF) using the line spread function (LSF) method. A novel plane source was prepared using thin layer chromatography (TLC) of a fluorine-18-fluorodeoxyglucose ((18)F-FDG) solution. The source was placed within a phantom, and imaged using the whole body (WB) two dimensional (2D) and three dimensional (3D) standard imaging protocols in a GE Discovery ST hybrid PET/CT scanner. Modulation transfer function was evaluated by determining the LSF, for various reconstruction methods and filters. The proposed MTF measurement method was validated against the conventional method, based on point spread function (PSF). Higher MTF values were obtained with 3D scanning protocol and 3D iterative reconstruction algorithm. All MTF obtained using 3D reconstruction algorithms showed better preservation of higher frequencies than the 2D algorithms. They also exhibited better contrast and resolution. MTF derived from LSF were more precise compared with those obtained from PSF since their reproducibility was better in all cases, providing a mean standard deviation of 0.0043, in contrary to the PSF method which gave 0.0405. In conclusion, the proposed method is novel and easy to implement for characterization of the signal transfer properties and image quality of PET/computed tomography (CT) systems. It provides an easy way to evaluate the frequency response of each kernel available. The proposed method requires cheap and easily accessible materials, available to the medical physicist in the nuclear medicine department. Furthermore, it is robust to aliasing and since this method is based on the LSF, is more resilient to noise due to greater data averaging than conventional PSF-integration techniques.