The proposal of SunCHem project is to produce methane from the hydrothermal liquefaction of microalgal biomass and recovery high-value products in a bio-refinery concept. In this sense the Environmental Biotechnology Laboratory of EPFL (LBE) has developing some techniques approaches and research to achieve higher biomass production and yield of bio-products as PUFA-3 and pigments. The aim of the present work was to study the effects of Intensity of Light Incident (ILI) and photoperiod (Photo) of a LED system illumination developed for test in flasks, evaluating the Scenedesmus vacuolatus (Chlorella fusca var. vacuolata) SAG 211 - 8B growth and chlorophyll content, using 22 experimental planning with 4 central points. The results obtained by Flow-cytometry analysis (Accuri C6 & CSampler) showed that in 120h time culture at the range applied for ILI (88 ≤ I ≤ 220 mol m-2 s-1) and Photo (8 ≤ h ≤ 16 h light) both independent variable influenced in the cell concentration and chlorophyll response. The highest cell concentration ( 0,214 g/L) was obtained at 220 mol m-2 s-1 and 16 h light. The correlation between events for biomass and chlorophyll content was verified at high biomass concentration. The results indicate that S. vacuolatus was able to grow in LED system illumination developed, suggesting an alternative for a large scale LED experiments at flasks level.