Efficient monitoring of organisms is at the foundation of protected area and biodiversity management. Such monitoring programs are based on a systematically selected set of survey locations that, while able to track trends at those locations through time, lack inference for the overall region being “monitored”. Advances in spatially-balanced sampling approaches offer alternatives but remain largely untested in marine ecosystems. This study evaluated the merit of using a two-stage, spatially-balanced survey framework, in conjunction with generalized additive models, to estimate epifauna cover at a reef-wide scale for mesophotic reefs within a large, cross-shelf marine park. Imagery acquired by an autonomous underwater vehicle was classified using a hierarchical scheme developed under the Collaborative and Automated Tools for Analysis of Marine Imagery (CATAMI). At a realistic image subsampling intensity, the two-stage, spatially-balanced framework provided accurate and precise estimates of reef-wide cover for a select number of epifaunal classes at the coarsest CATAMI levels, in particular bryozoan and porifera classes. However, at finer hierarchical levels, accuracy and/or precision of cover estimates declined, primarily because of the natural rarity of even the most common of these classes/morphospecies. Ranked predictor importance suggested that bathymetry, backscatter and derivative terrain variables calculated at their smallest analysis window scales (i.e. 81 m2) were generally the most important variables in the modeling of reef-wide cover. This study makes an important step in identifying the constraints and limitations that can be identified through a robust statistical approach to design and analysis. The two-stage, spatially-balanced framework has great potential for effective quantification of epifaunal cover in cross-shelf mesophotic reefs. However, greater image subsampling intensity than traditionally applied is required to ensure adequate observations for finer-level CATAMI classes and associated morphospecies.