The strengthening mechanism during aging treatment after 84.18% cold rolling of MP35N alloy was performed in this study. Mechanical properties were tested on universal tensile testing machine and the microstructures were analyzed by OM, SEM, TEM and EDS. The results show that the maximum micro-hardness, 704.26 HV, was obtained when MP35N alloy was aged at 500 ℃ for 4 h, at which state the tensile strength was 2641.16 MPa. The two values increased by 30.44% and 37.41% over cold rolling state respectively. Tear holes which formed during cold forming healed after aging and the TEM result revealed that the thin "twin cluster" consisting of 3 to 4 fine twins which thickness about 3 nm appeared after aged at 400 ℃. When the temperature increased to 500 ℃, twin size became larger, staggered and then "twin cluster" disappeared. The EDS results shows there is no obvious concentration of Mo atom occurred in the twins. Nano flake twins formed during aging treatment play a major role in strengthening.