The microstructural evolution of twinned martensite in water-quenched Fe-1.6 C (wt.%) alloys upon in situ heating was investigated using transmission electron microscopy (TEM). In the as-quenched samples, a high density of a body-centred cubic (bcc) {112} 〈111〉 -type twinning structure exists in the martensite structure. Upon in situ heating to approximately 200-250 °C, carbides (mainly θ-Fe3C cementite) accompanying a detwinning process were observed only in the originally twinned region. The carbides were absent in the originally untwinned (twin-free) region. The experimental results have suggested that the formation of the carbides depends on the twinning-boundary ω-Fe metastable phase, which can be stabilised by interstitial carbon atoms. When the specimens were heated, the twinning-boundary ω-Fe(C) transformed into carbide (mainly θ-Fe3C cementite) particles on the original {112} twinning planes. Further heating resulted in substantial recrystallisation of α-Fe fine particles, which formed immediately after martensite transformation. The results presented here should be helpful in understanding the microstructural evolution of various carbon steels.