The unicellular green microalgae, Haematococcus pluvialis, has been examined as a microbial source for the production of astaxanthin, which has been suggested as a food supplement for humans and is also prescribed as an ingredient in eye drops because of its powerful anti-oxidant properties. In this study, we estimated the effects of the slope of a V-shaped bottom design, the volumetric flow rate of air, height/diameter (H/D) ratio, and diameter of an air sparger on the performance of a photo-bioreactor. These parameters were selected because they are recognized as important factors effecting the mixing that produces increased cell density in the reactor. The mixing effect can be measured by changes in optical density in the bioreactor over a period of time. A 6 L indoor photo-bioreactor was prepared in a short time period of 24 h for the performance study. A bioreactor designed with a V-shaped bottom with a slope of 60° showed an optical density change of 0.052 at 680 nm, which was sixfold less than the change in a photo-bioreactor designed with a flat bottom. Studies exploring the effects of bioreactor configuration and a porous metal sparger with a 10 μm pore size showed the best performance at an H/D ratio of 6:1 and a sparger diameter of 1.3 cm, respectively. The optimal rate of air flow was 0.2 vvm. The indoor culture of microalgae in the photo-bioreactor was subsequently carried for an application study using the optimal values established for the important factors. The indoor culture system was composed of a light source controlled according to cell phase, a carbon dioxide feeder, a bag-type reactor with an H/D ratio of 6:1, and a temperature controller. Results demonstrated the efficient production of microalgal cells and astaxanthin in the amounts of 2.62 g/L and 78.37 mg/L, respectively, when using adequate hydrodynamic mixing. Furthermore, the optimal design of a photo-bioreactor can be applied for the phototropic culturing of other microalgae for mass production.