In this study, a combined simulation and experimental approach is utilized to investigate the influence of hatch spacing on the microstructure and as-built quality of 316L stainless steel (SS) samples fabricated by selective laser melting (SLM). A three-dimensional finite element model (FEM) is employed to investigate heat transfer and melt pool during the SLM of 316L SS. The phase transformation and variation of the thermo-physical properties of the materials are considered in this model. The effects of hatch spacing (H) on the temperature field, microstructure and melt pool size, overlap rate, surface quality, and relative density during the SLM of 316L SS are investigated. The simulated results indicate that, as the hatch spacing increases, the depth increases and the width of the melt pool decreases. Meanwhile, with the increase of hatch spacing, the simulated temperature of the subsequent tracks falls below the melting temperature of the first track. Moreover, the microstructures were found to coarsen with the increasing hatch spacing due to the reduced cooling rate. The optimized hatch spacing and overlap rate between adjacent tracks were obtained from numerical simulations. Simulation results illustrate that, when the optimized hatch spacing of 100 μm is adopted, fully dense parts with a smooth surface can be fabricated by SLM, thus experimentally validating the simulation results.