Alternative feeds are critical for the sustainable expansion of the marine finfish aquaculture industry. The industry uses wild-caught forage fish as a primary ingredient in farmed fish feeds. Alternative ingredients are needed to safeguard fisheries' sustainability and future aquaculture development. While there have been successes in alternative feeds, it is necessary to improve the existing options and identify alternative ingredients with higher concentrations of omega-3 polyunsaturated fatty acids (PUFAs). This study was designed to test a microalga, Schizochytrium sp., as a feed ingredient for sablefish (Anoplopoma fimbria) using six test diets. There were two fish-ingredient control diets: +FM+FO, which contained both fishmeal and fish oil, and −FM+FO, which contained fish oil, but no fishmeal. The remaining four diets contained alternative lipids and were completely fish-free. FF Flax contained flax oil as the only lipid source replacing fish oil. FF LowSc, FF ModSc, and FF HighSc contained a low, moderate, and high level of Schizochytrium sp. to replace fish oil, with flax oil content decreasing as the microalga increased. After a 20-week trial, sablefish growth differed across the feed treatments, with fish fed the high microalga-inclusion diet (FF HighSc) performing similarly to fish fed the fish-ingredient controls. Fulton's K condition factor, dry feed intake (DFI), and lipid productive value (LPV) were also influenced by treatment. For the four fish-free diets, specific growth rate increased with increasing inclusion of Schizochytrium sp. in the feed. Fillet fatty acid profiles were similarly influenced by diet treatment, generally reflecting the fatty acid profiles of the feed. Total fillet PUFAs were higher in sablefish from the fish-free treatments than the control treatments, with DHA increasing with increasing inclusion of dietary Schizochytrium. In contrast, EPA was higher in fillets from both fish-ingredient control treatments compared to fillets from the fish-free treatments, yet EPA remained higher than expected in sablefish fed the fish-free diets. Histologic evaluation of sablefish distal intestine and liver demonstrated that the microalga-inclusion diets were well tolerated and did not cause histomorphological changes in the tissues. These results suggest Schizochytrium sp. can increase PUFA concentrations in fish fillets without compromising fish health and growth, making it a viable ingredient for alternative sablefish feeds.