Non-neutrally buoyant suspension of bi-dispersed non-colloidal particles in viscous fluid rotating in a horizontal cylinder displays in-homogeneities in particle distribution with alternate bands of high and low particle concentrations along the symmetric axis of the cylinder. Experiments were carried out to characterize the axial segregation in bi-dispersed suspension at various filling fraction and rotation speed of cylinder. The mixture of same particles in absence of any suspending fluid did not show any segregation. However, in case of particles suspended in water it was observed that the rate of segregation increases with increase in filling fraction. Once the particles get segregated along the full length of the cylinder, these bands start to migrate along the tube axis finally merging to give wider bands. For a given filling fraction the rate of segregation increases with the angular speed of the rotating cylinder. When the tube is partially filled the particle segregation is observed at higher angular speed, whereas in fully filled case the segregation starts at much lower rotation speed for the same concentration of particles. The segregation pattern changes as the rotation speed is increased. At higher speed the centrifugal force dominates over gravitational and viscous drag forces and this result into completely different segregation patterns. We have also analyzed the evolution of concentration profile from the image analysis of the particles. (C) 2010 The Society of Powder Technology Japan. Published by Elsevier B.V. and The Society of Powder Technology Japan. All rights reserved.