Many marine organisms spend the early life history stages in neuston domain. Although the importance of ichthyoneuston, few studies were developed in the Southwest Atlantic Ocean. Therefore, this study aimed to improve the knowledge on ichthyoneuston of the Brazilian coast analyzing the vertical stratification and horizontal distribution of these organisms. Neuston samples were collected in daylight between 20°S and 23°S from February to April 2009 (late summer/early autumn; rainy season) and from August to September 2009 (late winter/early spring; dry season). Eggs of six taxa were identified: Anguilliformes, Engraulidae, Clupeidae, Synodontidae, Trichiuridae, and Maurolicus stehmanni (Sternoptychidae). The neustonic larval fish assemblage was composed by 40 families and 63 species. Mullidae and Myctophidae larvae were the most abundant in the rainy period while Mullidae and Mugilidae dominated in the dry season. Seasonal and spatial variation of larval fish assemblages in the neuston were structured by oceanographic features. The larval fish abundance on the outer slope stations may have been favored by the advection of an anticyclone that encompassed most of the study region during the cruise periods. In the rainy season, salinity and local depth structured the larval fish assemblage in a cross‐shelf gradient, while in the dry season the larval assemblage was structured around temperature and north‐south gradient. In the dry season, the cross‐shelf gradient was less pronounced mainly because of the low abundance and frequency of mesopelagic larvae. The low frequency and abundance of some species are probably related to the net avoidance of fish larvae during the day or dial vertical migration, as many species migrate to deeper layers during the day and ascend to neuston only at night. Nevertheless, the present study presented baseline information about the seasonal and spatial variation of the neustonic larval fish assemblage influenced by the oceanographic conditions in the Campos Basin. We recommend to additionally collect night samples to decrease larval escapement rates and to compare night versus day catches to further investigate the influence of daily migration in the neustonic larval fish in the area.