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Abstract: The diesel soot was extracted from the chamber of a running diesel engine using a total cylinder sampling system. A structural description and size distribution of the soot spherules were performed by means of high-resolution transmission electron microscopy (HRTEM). HRTEM image analysis was carried out to achieve semi-quantitative information about the soot organization. Different nanostructures were found in dependence upon the engine test conditions. A better organization of the graphitic layers occurred for the in-cylinder soot as the expansion stroke developed. The oxygen concentrations at sampling angles and the cylinder pressure were measured. The oxidation rates of the in-cylinder soot during the late combustion phase were calculated according to the differences of spherules size distributions between two consecutive sampling angles. The soot reactivity in dependence on the nanostructure was shown after deducting the effect of oxygen concentration and temperature. The increase of resistance toward oxidation was associated to the increase of the structural order for the in-cylinder soot. The results are a useful database for testing the structure–property relationship between the nanostructure and the reactivity of the in-cylinder diesel soot. [ABSTRACT FROM AUTHOR]