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Advanced characterisation techniques were used on LPBF Ti-6Al-4V samples produced on a heated base plate. When the substrate temperature is 100°C the elongation is 6\%, which increases and peaks at 10\% at 570°C, then sharply decreases to zero ductility at 770°C. At 100°C, a heavily strained and twinned microstructure, primarily composed of α+α', was observed and it was comparable to asbuilt microstructures obtained by conventional LPBF methods. At higher temperatures, twins are no longer present and instead nano-scale β precipitates are observed within α' and α, as well as dislocation networks (570°C) and tangles (770°C). Solute segregation at crystal defects was observed in all pre-heating conditions. Al and V segregation at microtwins was observed in the 100°C sample, reporting for the first time `selective' and mutually exclusive Al- and V-rich regions forming in adjacent twins. V segregation at dislocations was observed in the 570°C and 770°C samples, consistent with the higher preheating temperatures. High O contents were measured in all samples but with apparent opposing effects. At 100°C and 570°C was estimated to be below the critical threshold for O embrittlement and locally aids in maintaining a strength high by solid solution strengthening, whereas at 770°C it was above the threshold, therefore failing in a brittle fashion. Based on these observations, the initial increase in ductility from 100°C to 570°C is attributed to a reduction in microtwins and the dislocation networks acting as `soft barriers' for slip within a coarser microstructure. The lack of ductility at 770°C was attributed to local solute redistribution causing dislocation pinning and an increase of O content in this sample.