A comprehensive study on different physico-chemical properties of banana fiber is presented in this communication. This has led to a better understanding of its structure and function. Studies were carried out for both inner and outer region of inflorescence stem. FESEM studies clearly indicated that the inner-wall of the micro tubules consisted of closely spaced helical xylem fibers forming a cylindrical shape attached to it. XRD studies indicated that both the samples contained crystalline cellulose and high amount of amorphous content. The XPS peak located at ∼533 eV could be attributed to cellulose and hemicelluloses. The FTIR bands at ∼2925 cm−1 corresponded C-H stretching vibration in cellulose component. Fibers from outer and inner regions showed nearly similar cellulose content.