Mixotrophic cultivation of Haematococcus lacustris is one of the most promising strategies to produce natural astaxanthin. During mixotrophic growth, microalgae assimilate and metabolize organic carbon in addition to photosynthetic growth, resulting in increased biomass productivity. Several studies have evaluated the effect of different organic carbon sources on mixotrophic growth in various microalgae species. However, knowledge of detailed growth kinetics as a function of substrate concentration and light intensity is lacking. In this study, the growth kinetics of H. lacustris using four different carbon sources and the effect of light under mixotrophic and photoautotrophic conditions are described. Mixotrophic cultivation showed significant differences in respect to applied substrate and achieved maximum specific growth rates of 0.91 ± 0.13, 0.19 ± 0.05, 0.36 ± 0.05, and 0.23 ± 0.05 day−1, for acetate, methanol, glucose, and glycerol, respectively. Optimal growth at mixotrophic conditions using acetate was 1.8 times higher than the sum of hetero- and photoautotrophic growth. Furthermore, the optimum light intensity was 1.3 times higher for mixotrophic than for autotrophic growth. Thus, mixotrophy increases light intensity tolerance. These results indicate a strong interconnection between carbon metabolism and photosynthetic activity and lay the foundation for more detailed mathematical models describing the mixotrophic growth of H. lacustris.