An innovative in situ thermal approach in the domain of LPBF for Ti-6Al-4V fabrication has been carried out with results directing towards an improved fatigue life without the need for post-processing. The thermal process involves an additional laser scan with different process parameters to preheat the selected regions of each layer of the powder bed prior to their full melting. This preheating step influences the cooling rate, which in turn affects surface characteristics and subsurface microstructure, both of which are directly correlated with fatigue properties. A thorough analysis has been conducted by comparing the preheated samples with reference samples with no preheating. Without any additional thermal processing, the preheated samples showed a significant improvement over their reference counterparts. The optimized preheated sample showed an improved prior β-grain distribution with a circular morphology and thicker α laths within the even finer prior β-grain boundaries. Also, an overall increment of the c/a ratio of the HCP α has been observed, which yielded lattice strain relaxation in the localized grain structure. Furthermore, a less-profound surface roughness was observed in the preheated sample. The obtained microstructure with all these factors delivered a 10% improvement in its fatigue life with better mechanical strength overall.