Biofuels, fuels derived from biological materials, are considered an attractive alternative for fixing the ongoing energy crisis, due to their unlimited renewable resources and no net CO2 release into the atmosphere. The use of food crops such as corn, soybean, castor seeds, rapeseeds is undesirable due to the need for large cultivation areas, high loads on freshwater sources, and competition with food markets. Unicellular photosynthetic thallophytes, the microalgae, are being explored as a potential source of biodiesel, owing to their ease of cultivation, low nutrient requirements, little dependence on agricultural land, and ability to grow on waste water. Light-dependent growth is the most widely used technique for biodiesel production from microalgae. However, the inability to achieve a high biomass as a result of poor light penetration complicates the process as well as decreases the yield. Therefore, heterotrophic culture of microalgae in the dark when supplemented carbon source may be a way to overcome the drawbacks of autotrophic growth in mass culture. Strategies to increase the lipid content of the cells by manipulating their growth conditions are under study to increase lipid productivity from each harvest of batch cultures. Growth of microalgae in the stressful environment induced under low nitrogen conditions is known to cause algal cells to synthesize more lipids. Can other stresses increase lipid yields? The microalga, Chlorella kessleri, was grown under heterotrophic conditions with sucrose or lignocellulose hydrolysate as the carbon source. Lignocellulosic material is a readily available renewable source of carbon and its economic feasibility makes it an attractive carbon source. The nitrogen content in the media was manipulated at different concentrations ranging from 0 mM to 30 mM. With an increase in nitrogen deficiency, microalgae growth decreases, but the lipid content of the cells increases. The results from nitrogen starvation will help us to extrapolate to similar lipid increases with other starvation for nutrients such as phosphorus and iron. Therefore, heterotrophic growth of microalgae at different concentrations of phosphorus (0 mM-5 mM) and iron (0 ?M-20 ?M) is being studied by a cell counting method and chlorophyll measurements each day until stationary phase is reached. Furthermore, lipid content of cells at stationary phase is measured for each growth condition by Nile Red staining and a modified Bligh and Dyer method. Heterotrophic culture of microalgae with these novel strategies to increase the lipid content may become a sustainable alternative to petroleum.