CO 2 biofixation was investigated using tubular bioreactors (15 and 1.5 l) either in the presence of green algae Chlorella vulgaris or Nannochloropsis gaditana. The cultivation was carried out in the following conditions: temperature of 25 °C, inlet-CO 2 of 4 and 8 vol%, and artificial light enhancing photosynthesis. Higher biofixation were observed in 8 vol% CO 2 concentration for both microalgae cultures than in 4 vol%. Characteristic process parameters such as productivity, CO 2 fixation, and kinetic rate coefficient were determined and discussed. Simplified and advanced methods for determination of CO 2 fixation were compared. In a simplified method, it is assumed that 1 kg of produced biomass equals 1.88 kg recycled CO 2. Advance method is based on empirical results of the present study (formula with carbon content in biomass). It was observed that application of the simplified method can generate large errors, especially if the biomass contains a relatively low amount of carbon. N. gaditana is the recommended species for CO 2 removal due to a high biofixation rate—more than 1.7 g/l/day. On day 10 of cultivation, the cell concentration was more than 1.7 × 10 7 cells/ml. In the case of C. vulgaris, the maximal biofixation rate and cell concentration did not exceed 1.4 g/l/day and 1.3 × 10 7 cells/ml, respectively.