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In the present study, a commercial high pressure, common rail injection system for automotive DI diesel engines was fed with a conventional diesel fuel, a bio-derived fuel and a blend of them. The comparison of spray characteristics was carried out in terms of tip penetration and cone angles; the fuel spray, generated by rail pressures ranging from 60 MPa to 120 MPa, developed in an atmospheric chamber. The experimental set-up is based on a laser sheet technique. The radiation scattered by the spray, generated by a Nd-Yag pulsed laser, is collected by a CCD camera and fed to a frame grabber. A suitably set-up automatic image analysis process allows not only to determine the spray average development in terms of its geometric characteristics, but also to analyse in detail its internal structure. In particular, a suitable elaboration allowed the evaluation of the probability of presence in space of spray liquid fractions.