Lipid production using microalgae is challenging for producing low-value-added products. Harnessing microalgae for their fast and efficient CO₂ fixation capabilities may be more reasonable since algal biomass can be utilized as a precursor for various products in a biorefinery approach. This study aimed to optimize the productivity and efficiency of Microchloropsis salina biomass production in open thin-layer cascade (TLC) photobioreactors under physical simulation of suitable outdoor climate conditions, using an artificial seawater medium. Continuous operation proved to be the most suitable operating mode, allowing an average daily areal productivity of up to 27 g m⁻² d⁻¹ and CO₂ fixation efficiency of up to 100%. Process transfer from 8 m² to 50 m² TLC photobioreactors was demonstrated, but with reduced daily areal productivity of 21 g m⁻² d⁻¹ and a reduced CO₂ fixation efficiency, most probably due to increased temperatures at midday above 35 °C. An automated overnight switch-off of the circulation pumps was implemented successfully, reducing energy and freshwater requirements by ~40%. The ideal conditions for continuous production were determined to be a dilution rate of 0.150–0.225 d⁻¹, pH of 8.5, and total alkalinity of 200–400 ppm, facilitating efficient pilot-scale production of microalgal biomass in TLC photobioreactors.