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Use of a small tropical nursery in the Los Roques Archipelago by lemon sharks (Negaprion brevirostris) was investigated using acoustic telemetry. Twelve juvenile lemon sharks of three size classes were tracked between February 2014 and August 2015 in the Sebastopol Lagoon. Sharks were strongly site attached and remained in the lagoon for the duration of the study. Individuals in the smallest size class exhibited restricted movements within the innermost area of the lagoon in shallow water (< 1 m), over muddy substrate and along mangrove-lined shores. Sharks in the two larger size classes ranged further, in deeper water, over a wider range of substrates and more frequently near the lagoon entrance. Activity space varied among size classes, with home range (95% kernel utilization densities–KUD) of 0.42 km2 and core area (50% KUD) of 0.13 km2 for individuals in the smallest size class. For the medium and large size classes home ranges were 1.11 and 1.15 km2 and core areas were 0.33 and 0.35 km2 respectively. Space use as Minimum Convex Polygons differed among size classes, with overlap between the two largest size classes of 89%, compared with 40% between medium and smallest and 43% for largest and smallest size classes. Space use of lemon sharks in the Los Roques nursery illustrates variable use of habitat with varying environmental characteristics, likely reflecting a balance between predator avoidance and prey acquisition. Greater understanding of the use of nursery habitats for species such as lemon sharks, which use small, discrete nurseries over a broad geographical range can enhance our understanding of relationships between life history traits and environmental variability and management of populations. [ABSTRACT FROM AUTHOR]