This study present parameters effect on chitosan beads prepared via simultaneous crosslinking method. Chitosan macrospheres were prepared by precipitation method followed by crosslinking reaction with glutaraldehyde. Different parameters such as needle size, (31, 27.5,26, 25.5, 18 G), glutaraldehyde concentration (0.5, 1, 3, 5 wt%), concentration of chitosan (1, 2, 2.5, 3 wt%) and also concentration of NaOH (1, 2, 3 N) were used to obtained different size of chitosan beads. Increasing the diameter of the needle size will increase the particle size of chitosan beads. Increasing concentration of glutaraldehyde concentration will decreased the diameter of chitosan beads. Increasing the chitosan concentration will increase the diameter of the chitosan beads. Increasing NaOH concentration will decreasing the diameter of the chitosan beads The average particle size of the chitosan beads was obtained by calculating 100 diameters of it. The morphology and surface of the chitosan beads were characterized using Scanning Electron Microscope (SEM).