The southern flounder, Paralichthys lethostigma, is an important commercial and recreational marine flatfish that inhabits estuaries and shelf waters in the south Atlantic, from North Carolina through the Gulf coasts, with the exception of south Florida. Because juvenile and adult fish are highly euryhaline, it is a prime candidate for aquaculture. Methods for captive spawning of southern flounder are well developed; however, information on optimal culture requirements of the early larval stages is required for reliable mass production of juveniles. To determine the optimal photoperiod and salinity conditions for culture from hatching to day 15 post-hatching (d15ph), embryos were stocked into black 15-l tanks (75 l −1) under four photoperiods (24L:0D, 18L:6D, 12L:12D, and 6L:18D) and two salinities (25 and 34 ppt) in a 4×2 factorial design. Temperature was 18 °C, light intensity was 150 lx, and aeration was 50 ml min −1. Significant ( P<0.05) effects of photoperiod and salinity on growth (notochord length, wet and dry weights) were obtained. Growth increased with increasing photoperiod and salinity and was significantly greater at 24L and 18L than at 12L or 6L, and at 34 than at 25 ppt. On d11ph and d15ph, significant interactive effects between photoperiod and salinity on growth (wet and dry weights) were also evident. Growth of larvae reared at 25 ppt increased with increasing photoperiod to a maximum at 24L, while growth of larvae at 34 ppt reached a plateau at 18L. While there were no significant photoperiod effects on these parameters, larval survival, body water percentage, and larval osmolality on d15ph were significantly higher at 34 than at 25 ppt (41% vs. 16% survival; 322 vs. 288 mosM kg −1; and 84% vs. 76% water, respectively), suggesting stress and nonadaptation to 25 ppt, a salinity more nearly isoosmotic than full-strength seawater. Since larvae from both salinity treatments were neutrally or positively buoyant at 34 ppt, but negatively buoyant at 25 ppt, larvae reared at 25 ppt probably allocated energy to maintain vertical positioning, compromising growth and survival. The results demonstrate that growth and survival of early-stage southern flounder larvae are maximized under long photoperiods of 18–24L and in full-strength seawater. Longer photoperiods probably extend the time larvae have for feeding, while full-strength seawater salinity optimizes buoyancy and vertical positioning, conserving energy. The results show that early larval stage southern flounder larvae are not entirely euryhaline, which involves not only the ability to osmoregulate, but to conserve energy under reduced buoyancy. This is consistent with suboptimal vs. maximal growth of larvae reared at 25 and 34 ppt, respectively, under 18L (i.e., photoperiod×salinity interaction). This is also consistent with other reports that tolerance to lower salinities in these euryhaline flatfish increases post-metamorphosis when transition from a pelagic to benthic existence alleviates the need to counteract reduced buoyancy.