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For better understanding of soot formation and oxidation processes in a biodiesel spray flame, the morphology, microstructure and sizes of soot particles directly sampled in a spray flame fuelled with soy-methyl ester were investigated using transmission electron microscopy (TEM). The soot samples were taken at different axial locations in the spray flame, 40, 50 and 70 mm from injector nozzle, which correspond to soot formation, peak, and oxidation zones, respectively. The biodiesel spray flame was generated in a constant-volume combustion chamber under a diesel-like high pressure and temperature condition (6.7 MPa, 1000K). Density, diameter of primary particles and radius of gyration of soot aggregates reached a peak at 50 mm from the injector nozzle and was lower or smaller in the formation or oxidation zones of the spray.