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Different quantity and configurations of crystal defects were obtained in an austenite of Fe-30%Ni alloy and an ultra low carbon bainitic steel by different deformations and annealing times at high temperature. The boron segregation at grain boundaries and subgrain boundaries during air cooling were revealed by means of particle tracking autoradiography technique. It was found that the non-equilibrium segregation was resisted in the deformed grains after recovery and polygonization, the boron depletion was more in the recrystallized grains than in the deformed original grains during the cooling. The subgrain boundaries and polygonized dislocation cells had a significant effect on the boron non-equilibrium segregation during the air cooling, but the quantity of dispersed dislocations had not. The result indicated that during segregation process the interaction of boron atoms with dislocations was sensitive to the dislocations configuration rather than the total number of defects in grain.