In this experimental study the segregation behavior for fluidized mixtures of spherical and cylindrical particles is investigated. In industry, fluidization of particles featuring a wide range of shapes is common in various applications such as biomass gasification, drying applications, food processing and production of pharmaceuticals. Earlier publications have mainly focused on segregation of spherical particles of different volume or density. The particles used in this study have equal volume and density but a different shape. The main purpose of this work is to study de-mixing driven by particle shape. To analyze the particle distributions inside the fluidized bed, a Digital Image Analysis (DIA) technique has been developed, capable of capturing the particle positions and orientations within the bed over time. The experiments show that in the non-bubbling flow regime (at low fluidization velocities) rod-shaped particles may segregate, sinking to the bottom of the bed. In the bubbling flow regime (at higher fluidization velocities) segregation does not occur, because of bubble-induced mixing. Here strong alignment of the cylindrical particle's long axis with the flow is observed. The experimental results obtained give qualitative and quantitative insight in the behavior of non-spherical particles in fluidized beds and can be used for validation of numerical models concerning non-spherical particle mixing.