Directed energy deposition (DED) is a crucial branch of additive manufacturing (AM), performing repairs, cladding, and processing of multi-material components. 316L austenitic stainless steel is widely used in applications such as the food, aerospace, automotive, marine, energy, biomedical, and nuclear reactor industries. Nevertheless, there is need for process parameter optimization and a comprehensive understanding of the individual and complex synergistic effects of process parameters on the geometry, microstructure, and properties of the deposited material or component. This is essential for ensuring repeatable manufacturing of parts across a single or series of platforms over time, or for minimizing defects such as porosity. In this study, the response surface methodology (RSM) and central composite design (CCD) were employed to investigate the effects of laser power, laser scan speed, and powder mass flow rate on layer thickness, density, microstructure, and microhardness of 316L steel processed by Laser Engineered Net Shaping (LENS®) DED. Polynomial empirical prediction models correlating the applied processing parameters and the studied responses were developed.