The production of astaxanthin in the microalga Haematococcus pluvialis has been investigated using a sequential methodology based on the application of two types of statistical designs. The employed preliminary experiment was a fractional factorial design 2 6 in which the factors studied were: excessive irradiance and nitrate starvation, phosphate deficiency, acetate supplementation, salt stress, and elevated temperature. The experimental results indicate that the amount of astaxanthin accumulation in the cells can be enhanced by excessive irradiance and nitrate starvation whereas the other factors tested did not yield any enhancement. In the subsequent experiment, a central composite design was applied with four variables, light intensity, nitrate, phosphate, and acetate, at five levels each. The optimal conditions for the highest astaxanthin production were found to be 1040 E/(m 2 •s) light intensity, 0.04 g/L nitrate, 0.31 g/L phosphate, 0.05 g/L acetate concentration.