— The spray characteristic of the injected fuel is mainly depends upon fuel injection pressure, temperature, ambient pressure, fuel viscosity and fuel density. An experimental study was conducted to examine the effect of injection pressure on the spray was injected into direct injection (DI) diesel engine in the atmospheric condition. In Diesel engine, the window of 20 mm diameter hole and the transparent quartz glass materials were used for visualizing spray characteristics of combustion chamber at right angle triangle position. The varying Injection pressure of 180 - 240 bar and the engine was hand cranked for conducting the experiments. Spray characteristics for Jatropha oil methyl ester (JOME) and diesel were studied experimentally. Spray tip penetration and spray cone angle were measured in a combustion chamber of Direct Injection diesel engine by employing high speed Digital camera using Mie Scattering Technique and ImageJ software. The study shows the JOME gives longer spray tip penetration and smaller spray cone angle than those of diesel fuels. The Spray breakup region (Reynolds number, Weber number), Injection velocity and Sauter Mean Diameter (SMD) were determined for diesel and JOME. SMD decreases for JOME than diesel and the Injection velocity, Reynolds Number, Weber Number Increases for JOME than diesel.